U.S. patent application number 16/966961 was filed with the patent office on 2021-11-25 for tricyclic heterocycle compounds useful as hiv integrase inhibitors.
This patent application is currently assigned to Merck Sharp & Dohme Corp.. The applicant listed for this patent is Merck Sharp & Dohme Corp.. Invention is credited to Mark W. Embrey, Timothy John Hartingh, Marc Labroli, Izzat T. Raheem.
Application Number | 20210361662 16/966961 |
Document ID | / |
Family ID | 1000005809484 |
Filed Date | 2021-11-25 |
United States Patent
Application |
20210361662 |
Kind Code |
A1 |
Embrey; Mark W. ; et
al. |
November 25, 2021 |
TRICYCLIC HETEROCYCLE COMPOUNDS USEFUL AS HIV INTEGRASE
INHIBITORS
Abstract
The present invention relates to Tricyclic Heterocycle Compounds
of Formula (I): and pharmaceutically acceptable salts or prodrug
thereof, wherein X, Y, Z, R.sup.1 and n are as defined herein. The
present invention also relates to compositions comprising at least
one Tricyclic Heterocycle Compound, and methods of using the
Tricyclic Heterocycle Compounds for treating or preventing HIV
infection in a subject. ##STR00001##
Inventors: |
Embrey; Mark W.;
(Harleysville, PA) ; Hartingh; Timothy John;
(Richlandtown, PA) ; Labroli; Marc; (Moorestown,
NJ) ; Raheem; Izzat T.; (Doylestown, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Merck Sharp & Dohme Corp. |
Rahway |
NJ |
US |
|
|
Assignee: |
Merck Sharp & Dohme
Corp.
Rahway
NJ
|
Family ID: |
1000005809484 |
Appl. No.: |
16/966961 |
Filed: |
February 11, 2019 |
PCT Filed: |
February 11, 2019 |
PCT NO: |
PCT/US2019/017409 |
371 Date: |
August 3, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62630974 |
Feb 15, 2018 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/427 20130101;
A61K 31/5365 20130101; C07D 498/18 20130101; A61K 31/505 20130101;
A61K 31/635 20130101; A61K 31/7068 20130101; A61K 31/53 20130101;
C07D 471/18 20130101; A61K 31/675 20130101; A61K 31/513 20130101;
A61K 31/4418 20130101; A61K 31/52 20130101 |
International
Class: |
A61K 31/53 20060101
A61K031/53; C07D 471/18 20060101 C07D471/18; C07D 498/18 20060101
C07D498/18; A61K 31/513 20060101 A61K031/513; A61K 31/7068 20060101
A61K031/7068; A61K 31/427 20060101 A61K031/427; A61K 31/635
20060101 A61K031/635; A61K 31/4418 20060101 A61K031/4418; A61K
31/675 20060101 A61K031/675; A61K 31/505 20060101 A61K031/505; A61K
31/52 20060101 A61K031/52; A61K 31/5365 20060101 A61K031/5365 |
Claims
1. A compound of the formula: ##STR00118## or a pharmaceutically
acceptable salt thereof, wherein: X is --CR.sup.2R.sup.3, --O-- or
--NR.sup.4; Y is --CHR.sup.4--CR.sup.2R.sup.3, --CR.sup.2R.sup.3,
--O-- or --NR.sup.4; Z is --CR.sup.2R.sup.3--CHR.sup.4,
--CR.sup.2R.sup.3; each occurrence of R.sup.1 is independently
selected from the group consisting of halo, hydroxyl, C1-6 alkyl
and --O--(C1-C6 alkyl); each occurrence of R.sup.2 is independently
selected from the group consisting of hydrogen, halo, hydroxyl,
C1-6 alkyl and --O--(C1-C6 alkyl); each occurrence of R.sup.3 is
independently selected from the group consisting of hydrogen, halo,
hydroxyl, C1-6 alkyl and --O--(C1-C6 alkyl); each occurrence of
R.sup.4 is independently selected from the group consisting of
hydrogen or C1-6 alkyl; n is an integer between zero and three.
2. The compound of claim 1 wherein X is --CR.sup.2R.sup.3, or a
pharmaceutically acceptable salt thereof.
3. The compound of claim 1 wherein Y is
--CHR.sup.4--CR.sup.2R.sup.3, or a pharmaceutically acceptable salt
thereof.
4. The compound of claim 1 wherein Y is --CR.sup.2R.sup.3, or a
pharmaceutically acceptable salt thereof.
5. The compound of claim 1 wherein Y is --O--, or a
pharmaceutically acceptable salt thereof.
6. The compound of claim 1 wherein Y is --NR.sup.4, or a
pharmaceutically acceptable salt thereof.
7. The compound of claim 1 wherein Z is
--CR.sup.2R.sup.3--CHR.sup.4, or a pharmaceutically acceptable salt
thereof.
8. The compound of claim 1 wherein Z is --CR.sup.2R.sup.3, or a
pharmaceutically acceptable salt thereof.
9. The compound of claim 1 wherein each occurrence of R.sup.2 is
independently selected from the group consisting of hydrogen,
hydroxyl, and --OCH.sub.3, or a pharmaceutically acceptable salt
thereof.
10. The compound of claim 1 wherein R.sup.4 is hydrogen or methyl,
or a pharmaceutically acceptable salt thereof.
11. The compound of claim 1 wherein n is two or three, or a
pharmaceutically acceptable salt thereof.
12. The compound of claim 1 selected from: ##STR00119##
##STR00120## ##STR00121## or a pharmaceutically acceptable salt
thereof.
13. A pharmaceutical composition comprising an effective amount of
a compound according to claim 1, or a pharmaceutically acceptable
salt thereof, and a pharmaceutically acceptable carrier.
14. A method for the inhibition of HIV integrase in a subject in
need thereof which comprises administering to the subject an
effective amount of the compound according to claim 1, or a
pharmaceutically acceptable salt thereof.
15. A method for the treatment of infection by HIV or for the
treatment of AIDS in a subject in need thereof, which comprises
administering to the subject an effective amount of the compound
according to of claim 1, or a pharmaceutically acceptable salt
thereof.
16. The pharmaceutical composition of claim 13, further comprising
one or more additional therapeutic agents selected from,
raltegravir, lamivudine, abacavir, ritonavir, dolutegravir,
arunavir, atazanavir, emtricitabine, tenofovir, elvitegravir,
rilpivirine and lopinavir.
17. The method of claim 15, further comprising administering to the
subject one or more additional therapeutic agents selected from
raltegravir, lamivudine, abacavir, ritonavir, dolutegravir,
arunavir, atazanavir, emtricitabine, tenofovir, elvitegravir,
rilpivirine and lopinavir, wherein the amounts administered of the
compound of claim 1 and the one or more additional therapeutic
agents, are together effective to treat infection by HIV or to
treat, prevent or delay the onset or progression of AIDS.
18. A compound according to claim 1, or a pharmaceutically
acceptable salt thereof, for use in the preparation of a medicament
for the inhibition of HIV integrase, for the treatment or
prophylaxis of infection by HIV, or for the treatment, prophylaxis,
or delay in the onset or progression of AIDS in a subject in need
thereof.
19. (canceled)
Description
FIELD OF THE INVENTION
[0001] The present invention relates to Tricyclic Heterocycle
Compounds, compositions comprising at least one Tricyclic
Heterocycle Compound, and methods of using the Tricyclic
Heterocycle Compounds for treating or preventing HIV infection in a
subject.
BACKGROUND OF THE INVENTION
[0002] A retrovirus designated human immunodeficiency virus (HIV),
particularly the strains known as HIV type-1 (HIV-1) virus and
type-2 (HIV-2) virus, is the etiological agent of the complex
disease that includes progressive destruction of the immune system
(acquired immune deficiency syndrome; AIDS) and degeneration of the
central and peripheral nervous system. A common feature of
retrovirus replication is the insertion by virally-encoded
integrase of +proviral DNA into the host cell genome, a required
step in HIV replication in human T-lymphoid and monocytoid cells.
Integration is believed to be mediated by integrase in three steps:
assembly of a stable nucleoprotein complex with viral DNA
sequences; cleavage of two nucleotides from the 3' termini of the
linear proviral DNA, and covalent joining of the recessed 3' OH
termini of the proviral DNA at a staggered cut made at the host
target site. The fourth step in the process, repair synthesis of
the resultant gap, may be accomplished by cellular enzymes.
[0003] Nucleotide sequencing of HIV shows the presence of a pol
gene in one open reading frame [Rather, L. et al., Nature, 313,
277(1985)]. Amino acid sequence homology provides evidence that the
pol sequence encodes reverse transcriptase, integrase and an HIV
protease [Toh, H. et al., EMBO J. 4, 1267 (1985); Power, M. D. et
al., Science, 231, 1567 (1986); Pearl, L. H. et al., Nature, 329,
351 (1987)]. All three enzymes have been shown to be essential for
the replication of HIV.
[0004] It is known that some antiviral compounds which act as
inhibitors of HIV replication are effective agents in the treatment
of AIDS and similar diseases, including reverse transcriptase
inhibitors such as azidothymidine (AZT) and efavirenz and protease
inhibitors such as indinavir and nelfinavir. The compounds of this
invention are inhibitors of HIV integrase and inhibitors of HIV
replication.
SUMMARY OF THE INVENTION
[0005] In one aspect, the present invention provides Compounds of
Formula (I):
##STR00002##
or a pharmaceutically acceptable salt thereof, wherein:
[0006] X is --CR.sup.2R.sup.3, --O-- or --NR.sup.4;
[0007] Y is --CHR.sup.4--CR.sup.2R.sup.3, --CR.sup.2R.sup.3, --O--
or --NR.sup.4;
[0008] Z is --CR.sup.2R.sup.3--CHR.sup.4, --CR.sup.2R.sup.3;
[0009] each occurrence of R.sup.1 is independently selected from
the group consisting of halo, hydroxyl, C.sub.1-6 alkyl and
--O--(C.sub.1-C.sub.6 alkyl);
[0010] each occurrence of R.sup.2 is independently selected from
the group consisting of hydrogen, halo, hydroxyl, C.sub.1-6 alkyl
and --O--(C.sub.1-C.sub.6 alkyl);
[0011] each occurrence of R.sup.3 is independently selected from
the group consisting of hydrogen, halo, hydroxyl, C.sub.1-6 alkyl
and --O--(C.sub.1-C.sub.6 alkyl);
[0012] each occurrence of R.sup.4 is independently selected from
the group consisting of hydrogen or C.sub.1-6 alkyl;
[0013] n is an integer between zero and three.
[0014] The Compounds of Formula (I) (also referred to herein as the
"Tricyclic Heterocycle Compounds") and pharmaceutically acceptable
salts or prodrugs thereof may be useful, for example, for
inhibiting HIV viral replication or replicon activity, or for
treating or preventing HIV infection in a subject. Without being
bound by any specific theory, it is believed that the Tricyclic
Heterocycle Compounds inhibit HIV viral replication by inhibiting
HIV Integrase.
[0015] Accordingly, the present invention provides methods for
treating or preventing HIV infection in a subject, comprising
administering to the subject an effective amount of at least one
Tricyclic Heterocycle Compound.
[0016] The details of the invention are set forth in the
accompanying detailed description below.
[0017] Although any methods and materials similar to those
described herein may be used in the practice or testing of the
present invention, illustrative methods and materials are now
described. Other embodiments, aspects and features of the present
invention are either further described in or will be apparent from
the ensuing description, examples and appended claims.
DETAILED DESCRIPTION OF THE INVENTION
[0018] The present invention includes Tricyclic Heterocycle
Compounds, compositions comprising at least one Tricyclic
Heterocycle Compound, and methods of using the Tricyclic
Heterocycle Compounds for treating or preventing HIV infection in a
subject.
Definitions and Abbreviations
[0019] The terms used herein have their ordinary meaning and the
meaning of such terms is independent at each occurrence thereof.
That notwithstanding and except where stated otherwise, the
following definitions apply throughout the specification and
claims. Chemical names, common names, and chemical structures may
be used interchangeably to describe the same structure. These
definitions apply regardless of whether a term is used by itself or
in combination with other terms, unless otherwise indicated. Hence,
the definition of "alkyl" applies to "alkyl" as well as the "alkyl"
portions of "hydroxyalkyl," "haloalkyl," "--O-alkyl," etc.
[0020] As used herein, and throughout this disclosure, the
following terms, unless otherwise indicated, shall be understood to
have the following meanings:
[0021] A "subject" is a human or non-human mammal. In one
embodiment, a subject is a human. In another embodiment, a subject
is a primate. In another embodiment, a subject is a monkey. In
another embodiment, a subject is a chimpanzee. In still another
embodiment, a subject is a rhesus monkey.
[0022] The term "effective amount" as used herein, refers to an
amount of Tricyclic Heterocycle Compound and/or an additional
therapeutic agent, or a composition thereof that is effective in
inhibiting HIV replication and in producing the desired
therapeutic, ameliorative, inhibitory or preventative effect when
administered to a subject suffering from HIV infection or AIDS. In
the combination therapies of the present invention, an effective
amount can refer to each individual agent or to the combination as
a whole, wherein the amounts of all agents administered are
together effective, but wherein the component agent of the
combination may not be present individually in an effective
amount.
[0023] The terms "treating" or "treatment" as used herein with
respect to an HIV viral infection or AIDS, includes inhibiting the
severity of HIV infection or AIDS a disease, i.e., arresting or
reducing the development of the HIV infection or AIDS a disease or
its clinical symptoms; or relieving the HIV infection or AIDS a
disease, i.e., causing regression of the severity of HIV infection
or AIDS a disease or its clinical symptoms.
[0024] The terms "preventing," or "prohylaxis," as used herein with
respect to an HIV viral infection or AIDS, refers to reducing the
likelihood or severity of HIV infection or AIDS.
[0025] The term "alkyl," as used herein, refers to an aliphatic
hydrocarbon group having one of its hydrogen atoms replaced with a
bond. An alkyl group may be straight or branched and contain from
about 1 to about 20 carbon atoms. In one embodiment, an alkyl group
contains from about 1 to about 12 carbon atoms. In different
embodiments, an alkyl group contains from 1 to 6 carbon atoms
(C.sub.1-C.sub.6 alkyl) or from about 1 to about 4 carbon atoms
(C.sub.1-C.sub.4 alkyl). Non-limiting examples of alkyl groups
include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl,
isobutyl, tert-butyl, n-pentyl, neopentyl, isopentyl, n-hexyl,
isohexyl and neohexyl. In one embodiment, an alkyl group is linear.
In another embodiment, an alkyl group is branched. Unless otherwise
indicated, an alkyl group is unsubstituted.
[0026] The term "halo," as used herein, means --F, --Cl, --Br or
--I.
[0027] The term "haloalkyl," as used herein, refers to an alkyl
group as defined above, wherein one or more of the alkyl group's
hydrogen atoms has been replaced with a halogen. In one embodiment,
a haloalkyl group has from 1 to 6 carbon atoms. In another
embodiment, a haloalkyl group is substituted with from 1 to 3 F
atoms. Non-limiting examples of haloalkyl groups include
--CH.sub.2F, --CHF.sub.2, --CF.sub.3, --CH.sub.2Cl and --CCl.sub.3.
The term "C.sub.1-C.sub.6 haloalkyl" refers to a haloalkyl group
having from 1 to 6 carbon atoms.
[0028] The term "substituted" means that one or more hydrogens on
the designated atom is replaced with a selection from the indicated
group, provided that the designated atom's normal valency under the
existing circumstances is not exceeded, and that the substitution
results in a stable compound. Combinations of substituents and/or
variables are permissible only if such combinations result in
stable compounds. By "stable compound" or "stable structure" is
meant a compound that is sufficiently robust to survive isolation
to a useful degree of purity from a reaction mixture, and
formulation into an efficacious therapeutic agent.
[0029] The term "in substantially purified form," as used herein,
refers to the physical state of a compound after the compound is
isolated from a synthetic process (e.g., from a reaction mixture),
a natural source, or a combination thereof. The term "in
substantially purified form," also refers to the physical state of
a compound after the compound is obtained from a purification
process or processes described herein or well-known to the skilled
artisan (e.g., chromatography, recrystallization and the like), in
sufficient purity to be characterizable by standard analytical
techniques described herein or well-known to the skilled
artisan.
[0030] It should also be noted that any carbon as well as
heteroatom with unsatisfied valences in the text, schemes, examples
and tables herein is assumed to have the sufficient number of
hydrogen atom(s) to satisfy the valences.
[0031] When a functional group in a compound is termed "protected",
this means that the group is in modified form to preclude undesired
side reactions at the protected site when the compound is subjected
to a reaction. Suitable protecting groups will be recognized by
those with ordinary skill in the art as well as by reference to
standard textbooks such as, for example, T. W. Greene et al,
Protective Groups in Organic Synthesis (1991), Wiley, New York.
[0032] When any substituent or variable (e.g., R.sup.2 and R.sup.3)
occurs more than one time in any constituent or in Formula (I), its
definition on each occurrence is independent of its definition at
every other occurrence, unless otherwise indicated.
[0033] As used herein, the term "composition" is intended to
encompass a product comprising the specified ingredients in the
specified amounts, as well as any product which results from
combination of the specified ingredients in the specified
amounts.
[0034] Prodrugs and solvates of the compounds of the invention are
also contemplated herein. A discussion of prodrugs is provided in
T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems
(1987) 14 of the A.C.S. Symposium Series, and in Bioreversible
Carriers in Drug Design, (1987) Edward B. Roche, ed., American
Pharmaceutical Association and Pergamon Press. The term "prodrug"
means a compound (e.g., a drug precursor) that is transformed in
vivo to provide a Tricyclic Heterocycle Compound or a
pharmaceutically acceptable salt of the compound. The
transformation may occur by various mechanisms (e.g., by metabolic
or chemical processes), such as, for example, through hydrolysis in
blood. For example, if a Tricyclic Heterocycle Compound or a
pharmaceutically acceptable salt, hydrate or solvate of the
compound contains a carboxylic acid functional group, a prodrug can
comprise an ester formed by the replacement of the hydrogen atom of
the acid group with a group such as, for example,
(C.sub.1-C.sub.8)alkyl, (C.sub.2-C.sub.12)alkanoyloxymethyl,
1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms,
1-methyl-1-(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms,
alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms,
1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms,
1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon
atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon
atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon
atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,
di-N,N--(C.sub.1-C.sub.2)alkylamino(C.sub.2-C.sub.3)alkyl (such as
(3-dimethylaminoethyl), carbamoyl-(C.sub.1-C.sub.2)alkyl, N,N-di
(C.sub.1-C.sub.2)alkylcarbamoyl-(C.sub.1-C.sub.2)alkyl and
piperidino-, pyrrolidino- or morpholino(C.sub.2-C.sub.3)alkyl, and
the like.
[0035] Similarly, if a Tricyclic Heterocycle Compound contains an
alcohol functional group, a prodrug can be formed by the
replacement of one or more of the hydrogen atoms of the alcohol
groups with a group such as, for example,
(C.sub.1-C.sub.6)alkanoyloxymethyl, C.sub.6)alkanoyloxy)ethyl,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
(C.sub.1-C.sub.6)alkoxycarbonyloxymethyl,
N--(C.sub.1-C.sub.6)alkoxycarbonylaminomethyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkyl,
.alpha.-amino(C.sub.1-C.sub.4)alkylene-aryl, arylacyl and
.alpha.-aminoacyl, or .alpha.-aminoacyl-.alpha.-aminoacyl, where
each .alpha.-aminoacyl group is independently selected from the
naturally occurring L-amino acids, or glycosyl (the radical
resulting from the removal of a hydroxyl group of the hemiacetal
form of a carbohydrate).
[0036] If a Tricyclic Heterocycle Compound incorporates an amine
functional group, a prodrug can be formed by the replacement of a
hydrogen atom in the amine group with a group such as, for example,
R-carbonyl-, RO-carbonyl-, NRR'-carbonyl- wherein R and R' are each
independently (C.sub.1-C.sub.10)alkyl, (C.sub.3-C.sub.7)
cycloalkyl, benzyl, a natural .alpha.-aminoacyl,
--C(OH)C(O)OY.sup.1 wherein Y.sup.1 is H, (C.sub.1-C.sub.6)alkyl or
benzyl, --C(OY.sup.2)Y.sup.3 wherein Y.sup.2 is (C.sub.1-C.sub.4)
alkyl and Y.sup.3 is (C.sub.1-C.sub.6)alkyl; carboxy
(C.sub.1-C.sub.6)alkyl; amino(C.sub.1-C.sub.4)alkyl or mono-N- or
di-N,N--(C.sub.1-C.sub.6)alkylaminoalkyl; --C(Y.sup.4)Y.sup.5
wherein Y.sup.4 is H or methyl and Y.sup.5 is mono-N- or
di-N,N--(C.sub.1-C.sub.6)alkylamino morpholino; piperidin-1-yl or
pyrrolidin-1-yl, and the like.
[0037] Pharmaceutically acceptable esters of the present compounds
include the following groups: (1) carboxylic acid esters obtained
by esterification of the hydroxy group of a hydroxyl compound, in
which the non-carbonyl moiety of the carboxylic acid portion of the
ester grouping is selected from straight or branched chain alkyl
(e.g., methyl, ethyl, n-propyl, isopropyl, t-butyl, sec-butyl or
n-butyl), alkoxyalkyl (e.g., methoxymethyl), aralkyl (e.g.,
benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (e.g.,
phenyl optionally substituted with, for example, halogen,
C.sub.1-4alkyl, --O--(C.sub.1-4alkyl) or amino); (2) sulfonate
esters, such as alkyl- or aralkylsulfonyl (for example,
methanesulfonyl); (3) amino acid esters, including those
corresponding to both natural and non-natural amino acids (e.g.,
L-valyl or L-isoleucyl); (4) phosphonate esters and (5) mono-, di-
or triphosphate esters. The phosphate esters may be further
esterified by, for example, a C.sub.1-20 alcohol or reactive
derivative thereof, or by a 2,3-di (C.sub.6-24)acyl glycerol.
[0038] One or more compounds of the invention may exist in
unsolvated as well as solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, and the like, and it is
intended that the invention embrace both solvated and unsolvated
forms. "Solvate" means a physical association of a compound of this
invention with one or more solvent molecules. This physical
association involves varying degrees of ionic and covalent bonding,
including hydrogen bonding. In certain instances the solvate will
be capable of isolation, for example when one or more solvent
molecules are incorporated in the crystal lattice of the
crystalline solid. "Solvate" encompasses both solution-phase and
isolatable solvates. Non-limiting examples of solvates include
ethanolates, methanolates, and the like. A "hydrate" is a solvate
wherein the solvent molecule is water.
[0039] One or more compounds of the invention may optionally be
converted to a solvate. Preparation of solvates is generally known.
Thus, for example, M. Caira et al, J. Pharmaceutical Sci., 93(3),
601-611 (2004) describe the preparation of the solvates of the
antifungal fluconazole in ethyl acetate as well as from water.
Similar preparations of solvates, hemisolvates, hydrates and the
like are described by E. C. van Tonder et al, AAPS PharmSciTech.
5(1), article 12 (2004); and A. L. Bingham et al, Chem. Commun.,
603-604 (2001). A typical, non-limiting, process involves
dissolving the inventive compound in desired amounts of the desired
solvent (organic or water or mixtures thereof) at a higher than
room temperature, and cooling the solution at a rate sufficient to
form crystals which are then isolated by standard methods.
Analytical techniques such as, for example IR spectroscopy, show
the presence of the solvent (or water) in the crystals as a solvate
(or hydrate).
[0040] The Tricyclic Heterocycle Compounds can form salts which are
also within the scope of this invention. Reference to a Tricyclic
Heterocycle Compound herein is understood to include reference to
salts thereof, unless otherwise indicated. The term "salt(s)", as
employed herein, denotes acidic salts formed with inorganic and/or
organic acids, as well as basic salts formed with inorganic and/or
organic bases. In addition, when a Tricyclic Heterocycle Compound
contains both a basic moiety, such as, but not limited to a
pyridine or imidazole, and an acidic moiety, such as, but not
limited to a carboxylic acid, zwitterions ("inner salts") may be
formed and are included within the term "salt(s)" as used herein.
In one embodiment, the salt is a pharmaceutically acceptable (i.e.,
non-toxic, physiologically acceptable) salt. In another embodiment,
the salt is other than a pharmaceutically acceptable salt. Salts of
the Compounds of Formula (I) may be formed, for example, by
reacting a Tricyclic Heterocycle Compound with an amount of acid or
base, such as an equivalent amount, in a medium such as one in
which the salt precipitates or in an aqueous medium followed by
lyophilization.
[0041] Exemplary acid addition salts include acetates, ascorbates,
benzoates, benzenesulfonates, bisulfates, borates, butyrates,
citrates, camphorates, camphorsulfonates, fumarates,
hydrochlorides, hydrobromides, hydroiodides, lactates, maleates,
methanesulfonates, naphthalenesulfonates, nitrates, oxalates,
phosphates, propionates, salicylates, succinates, sulfates,
tartarates, thiocyanates, toluenesulfonates (also known as
tosylates) and the like. Additionally, acids which are generally
considered suitable for the formation of pharmaceutically useful
salts from basic pharmaceutical compounds are discussed, for
example, by P. Stahl et al, Camille G. (eds.) Handbook of
Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich:
Wiley-VCH; S. Berge et al, Journal of Pharmaceutical Sciences
(1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics
(1986) 33 201-217; Anderson et al, The Practice of Medicinal
Chemistry (1996), Academic Press, New York; and in The Orange Book
(Food & Drug Administration, Washington, D.C. on their
website). These disclosures are incorporated herein by reference
thereto.
[0042] Exemplary basic salts include ammonium salts, alkali metal
salts such as sodium, lithium, and potassium salts, alkaline earth
metal salts such as calcium and magnesium salts, salts with organic
bases (for example, organic amines) such as dicyclohexylamine,
t-butyl amine, choline, and salts with amino acids such as
arginine, lysine and the like. Basic nitrogen-containing groups may
be quarternized with agents such as lower alkyl halides (e.g.,
methyl, ethyl, and butyl chlorides, bromides and iodides), dialkyl
sulfates (e.g., dimethyl, diethyl, and dibutyl sulfates), long
chain halides (e.g., decyl, lauryl, and stearyl chlorides, bromides
and iodides), arylalkyl halides (e.g., benzyl and phenethyl
bromides), and others.
[0043] All such acid salts and base salts are intended to be
pharmaceutically acceptable salts within the scope of the invention
and all acid and base salts are considered equivalent to the free
forms of the corresponding compounds for purposes of the
invention.
[0044] Diastereomeric mixtures can be separated into their
individual diastereomers on the basis of their physical chemical
differences by methods well-known to those skilled in the art, such
as, for example, by chromatography and/or fractional
crystallization. Enantiomers can be separated by converting the
enantiomeric mixture into a diastereomeric mixture by reaction with
an appropriate optically active compound (e.g., chiral auxiliary
such as a chiral alcohol or Mosher's acid chloride), separating the
diastereomers and converting (e.g., hydrolyzing) the individual
diastereomers to the corresponding pure enantiomers.
Sterochemically pure compounds may also be prepared by using chiral
starting materials or by employing salt resolution techniques.
Also, some of the Tricyclic Heterocycle Compounds may be
atropisomers (e.g., substituted biaryls) and are considered as part
of this invention. Enantiomers can also be directly separated using
chiral chromatographic techniques.
[0045] It is also possible that the Tricyclic Heterocycle Compounds
may exist in different tautomeric forms, and all such forms are
embraced within the scope of the invention. For example, all
keto-enol and imine-enamine forms of the compounds are included in
the invention.
[0046] Unless otherwise indicated, all stereoisomers (for example,
geometric isomers, optical isomers and the like) of the present
compounds (including those of the salts, solvates, hydrates, esters
and prodrugs of the compounds as well as the salts, solvates and
esters of the prodrugs), such as those which may exist due to
asymmetric carbons on various substituents, including enantiomeric
forms (which may exist even in the absence of asymmetric carbons),
rotameric forms, atropisomers, and diastereomeric forms, are
contemplated within the scope of this invention. If a Tricyclic
Heterocycle Compound incorporates a double bond or a fused ring,
both the cis- and trans-forms, as well as mixtures, are embraced
within the scope of the invention.
[0047] When a substituent on a chiral carbon atom is depicted
without stereospecificity (by using a straight line bond to a
chiral center), it is to be understood that both the alpha and beta
configurations of said substituent group are to be considered part
of the present invention. For example, the compound of the present
invention, which is drawn as follows:
##STR00003##
is understood to encompass both stereoisomers at the indicated
chiral center, the structures of which are as follows:
##STR00004##
[0048] In the Examples section below, compounds of the present
invention that have been purified as individual stereoisomers are
sometimes depicted in non-stereospecific form but identified using
one or more of the terms: "diastereomer 1," "diastereomer 2,"
"enantiomer A" and "enantiomer B." In this instance, the absolute
stereochemistry of each isolated diastereomer and enantiomeric
center has not been determined and the terms used above are used to
represent each individual purified stereochemicacally pure
compound.
[0049] Individual stereoisomers of the compounds of the invention
may, for example, be substantially free of other isomers, or may be
admixed, for example, as racemates or with all other, or other
selected, stereoisomers. The chiral centers of the present
invention can have the S or R configuration as defined by the IUPAC
1974 Recommendations. The use of the terms "salt", "solvate",
"ester", "prodrug" and the like, is intended to apply equally to
the salt, solvate, ester and prodrug of enantiomers, stereoisomers,
rotamers, tautomers, racemates or prodrugs of the inventive
compounds.
[0050] In the Compounds of Formula (I), the atoms may exhibit their
natural isotopic abundances, or one or more of the atoms may be
artificially enriched in a particular isotope having the same
atomic number, but an atomic mass or mass number different from the
atomic mass or mass number predominantly found in nature. The
present invention is meant to include all suitable isotopic
variations of the compounds of generic Formula I. For example,
different isotopic forms of hydrogen (H) include protium (.sup.1H)
and deuterium (.sup.2H). Protium is the predominant hydrogen
isotope found in nature. Enriching for deuterium may provide
certain therapeutic advantages, such as increasing in vivo
half-life or reducing dosage requirements, or may provide a
compound useful as a standard for characterization of biological
samples. Isotopically-enriched Compounds of Formula (I) can be
prepared without undue experimentation by conventional techniques
well known to those skilled in the art or by processes analogous to
those described in the Schemes and Examples herein using
appropriate isotopically-enriched reagents and/or intermediates. In
one embodiment, a Compound of Formula (I) has one or more of its
hydrogen atoms replaced with deuterium.
[0051] The Tricyclic Heterocycle Compounds may be useful in human
and veterinary medicine for treating or preventing HIV infection in
a subject. In one embodiment, the Tricyclic Heterocycle Compounds
can be inhibitors of HIV viral replication. In a specific
embodiment, the Tricyclic Heterocycle Compounds are inhibitors of
HIV-1. Accordingly, the Tricyclic Heterocycle Compounds may be
useful for treating HIV infections and AIDS. In accordance with the
invention, the Tricyclic Heterocycle Compounds can be administered
to a subject in need of treatment or prevention of HIV
infection.
[0052] Accordingly, in one embodiment, the invention provides
methods for treating HIV infection in a subject comprising
administering to the subject an effective amount of at least one
Tricyclic Heterocycle Compound or a pharmaceutically acceptable
salt thereof. In a specific embodiment, the present invention
provides methods for treating AIDS in a subject comprising
administering to the subject an effective amount of at least one
Tricyclic Heterocycle Compound or a pharmaceutically acceptable
salt thereof.
The Compounds of Formula (I)
[0053] The present invention provides Tricyclic Heterocycle
Compounds of Formula (I):
##STR00005##
and pharmaceutically acceptable salts thereof, wherein:
[0054] X is --CR.sup.2R.sup.3, --O-- or --NR.sup.4;
[0055] Y is --CHR.sup.4--CR.sup.2R.sup.3, --CR.sup.2R.sup.3, --O--
or --NR.sup.4;
[0056] Z is --CR.sup.2R.sup.3--CHR.sup.4, --CR.sup.2R.sup.3;
[0057] each occurrence of R.sup.1 is independently selected from
the group consisting of halo, hydroxyl, C.sub.1-6 alkyl and
--O--(C.sub.1-C.sub.6 alkyl);
[0058] each occurrence of R.sup.2 is independently selected from
the group consisting of hydrogen, halo, hydroxyl, C.sub.1-6 alkyl
and --O--(C.sub.1-C.sub.6 alkyl);
[0059] each occurrence of R.sup.3 is independently selected from
the group consisting of hydrogen, halo, hydroxyl, C.sub.1-6 alkyl
and --O--(C.sub.1-C.sub.6 alkyl);
[0060] each occurrence of R.sup.4 is independently selected from
the group consisting of hydrogen or C.sub.1-6 alkyl;
[0061] n is an integer between zero and three.
[0062] In an embodiment of the invention, X is --CR.sup.2R.sup.3.
In a class of the embodiment, X is CH.sub.2.
[0063] In an embodiment of the invention, Y is
--CHR.sup.4--CR.sup.2R.sup.3. In a class of the embodiment, Y is
--CH.sub.2CH.sub.2. In another embodiment, Y is --CR.sup.2R.sup.3.
In a class of the embodiment, Y is --CHOCH.sub.3. In another class
of the embodiment, Y is --CHOH. In another class of the embodiment,
Y is --CH.sub.2. In another class of the embodiment, Y is
--CF.sub.2. In another embodiment, Y is --O--. In another
embodiment, Y is --NR.sup.4. In a class of the embodiment, Y is
--NCH.sub.3.
[0064] In an embodiment of the invention, Z is
--CR.sup.2R.sup.3--CHR.sup.4. In a class of the embodiment, Z is
--CH.sub.2CH.sub.2. In another embodiment, Z is --CR.sup.2R.sup.3.
In a class of the embodiment, Z is --CH.sub.2.
[0065] In an embodiment of the invention, each occurrence of
R.sup.1 is halo.
[0066] In an embodiment of the invention, each occurrence of
R.sup.2 is independently selected from the group consisting of
hydrogen, halo, hydroxyl, and --OCH.sub.3.
[0067] In an embodiment of the invention, R.sup.4 is hydrogen or
methyl.
[0068] In an embodiment of the invention, n is two. In another
embodiment, n is three.
[0069] In one embodiment, variables X, Y, Z, R.sup.1 and n for the
Compounds of Formula (I) are selected independently of each
other.
[0070] In another embodiment, the Compounds of Formula (I) are in
substantially purified form.
[0071] It is to be understood that any of the aforementioned
embodiments may be combined with one or more separate
embodiments.
[0072] Other embodiments of the present invention include the
following:
[0073] (a) A pharmaceutical composition comprising an effective
amount of a Compound of Formula (I), and a pharmaceutically
acceptable carrier.
[0074] (b) The pharmaceutical composition of (a), further
comprising a second therapeutic agent selected from the group
consisting of HIV antiviral agents, immunomodulators, and
anti-infective agents.
[0075] (c) The pharmaceutical composition of (b), wherein the HIV
antiviral agent is an antiviral selected from the group consisting
of HIV protease inhibitors and HIV NNRTI inhibitors.
[0076] (d) A pharmaceutical combination that is (i) a Compound of
Formula (I) and (ii) a second therapeutic agent selected from the
group consisting of HIV antiviral agents, immunomodulators, and
anti-infective agents; wherein the Compound of Formula (I) and the
second therapeutic agent are each employed in an amount that
renders the combination effective for inhibiting HIV replication,
or for treating HIV infection and/or reducing the likelihood or
severity of symptoms of HIV infection.
[0077] (e) The combination of (d), wherein the HIV antiviral agent
is an antiviral selected from the group consisting of HIV protease
inhibitors and HIV NNRTI inhibitors.
[0078] (f) A method of inhibiting HIV replication in a subject in
need thereof which comprises administering to the subject an
effective amount of a Compound of Formula (I).
[0079] (g) A method of treating HIV infection and/or reducing the
likelihood or severity of symptoms of HIV infection in a subject in
need thereof which comprises administering to the subject an
effective amount of a Compound of Formula (I).
[0080] (h) The method of (g), wherein the Compound of Formula (I)
is administered in combination with an effective amount of at least
one second therapeutic agent selected from the group consisting of
HIV antiviral agents, immunomodulators, and anti-infective
agents.
[0081] (i) The method of (h), wherein the HIV antiviral agent is an
antiviral selected from the group consisting of HIV protease
inhibitors and HIV NNRTI inhibitors.
[0082] (j) A method of inhibiting HIV replication in a subject in
need thereof which comprises administering to the subject the
pharmaceutical composition of (a), (b) or (c) or the combination of
(d) or (e).
[0083] (k) A method of treating HIV infection and/or reducing the
likelihood or severity of symptoms of HIV infection in a subject in
need thereof which comprises administering to the subject the
pharmaceutical composition of (a), (b) or (c) or the combination of
(d) or (e).
[0084] Additional embodiments of the present invention include the
following:
[0085] (l) A pharmaceutical composition comprising an effective
amount of a pharmaceutically acceptable salt of a Compound of
Formula (I), and a pharmaceutically acceptable carrier.
[0086] (m) The pharmaceutical composition of (l), further
comprising a second therapeutic agent selected from the group
consisting of HIV antiviral agents, immunomodulators, and
anti-infective agents.
[0087] (n) The pharmaceutical composition of (m), wherein the HIV
antiviral agent is an antiviral selected from the group consisting
of HIV protease inhibitors and HIV NNRTI inhibitors.
[0088] (o) A pharmaceutical combination that is (i) a
pharmaceutically acceptable salt of a Compound of Formula (I) and
(ii) a second therapeutic agent selected from the group consisting
of HIV antiviral agents, immunomodulators, and anti-infective
agents; wherein the pharmaceutically acceptable salt of the
Compound of Formula (I) and the second therapeutic agent are each
employed in an amount that renders the combination effective for
inhibiting HIV replication, or for treating HIV infection and/or
reducing the likelihood or severity of symptoms of HIV
infection.
[0089] (p) The combination of (o), wherein the HIV antiviral agent
is an antiviral selected from the group consisting of HIV protease
inhibitors and HIV NNRTI inhibitors.
[0090] (q) A method of inhibiting HIV replication in a subject in
need thereof which comprises administering to the subject an
effective amount of a pharmaceutically acceptable salt of a
Compound of Formula (I).
[0091] (r) A method of treating HIV infection and/or reducing the
likelihood or severity of symptoms of HIV infection in a subject in
need thereof which comprises administering to the subject an
effective amount of a pharmaceutically acceptable salt of a
Compound of Formula (I).
[0092] (s) The method of (r), wherein the pharmaceutically
acceptable salt of the Compound of Formula (I) is administered in
combination with an effective amount of at least one second
therapeutic agent selected from the group consisting of HIV
antiviral agents, immunomodulators, and anti-infective agents.
[0093] (t) The method of (s), wherein the HIV antiviral agent is an
antiviral selected from the group consisting of HIV protease
inhibitors and HIV NS5B polymerase inhibitors.
[0094] (u) A method of inhibiting HIV replication in a subject in
need thereof which comprises administering to the subject the
pharmaceutical composition of (1), (m) or (n) or the combination of
(o) or (p).
[0095] (v) A method of treating HIV infection and/or reducing the
likelihood or severity of symptoms of HIV infection in a subject in
need thereof which comprises administering to the subject the
pharmaceutical composition of (1), (m) or (n) or the combination of
(o) or (p).
[0096] Further embodiments of the present invention include the
following:
[0097] (w) A pharmaceutical composition comprising an effective
amount of a Compound of Formula (I) or a pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable
carrier.
[0098] (x) The pharmaceutical composition of (w), further
comprising a second therapeutic agent selected from the group
consisting of HIV antiviral agents, immunomodulators, and
anti-infective agents.
[0099] (y) The pharmaceutical composition of (x), wherein the HIV
antiviral agent is an antiviral selected from the group consisting
of HIV protease inhibitors and HIV NNRTI inhibitors.
[0100] (z) A pharmaceutical combination that is (i) a Compound of
Formula (I) and (ii) or a pharmaceutically acceptable salt thereof,
a second therapeutic agent selected from the group consisting of
HIV antiviral agents, immunomodulators, and anti-infective agents;
wherein the Compound of Formula (I) and the second therapeutic
agent are each employed in an amount that renders the combination
effective for inhibiting HIV replication, or for treating HIV
infection and/or reducing the likelihood or severity of symptoms of
HIV infection.
[0101] (aa) The combination of (z), wherein the HIV antiviral agent
is an antiviral selected from the group consisting of HIV protease
inhibitors and HIV NNRTI inhibitors.
[0102] (bb) A method of inhibiting HIV replication in a subject in
need thereof which comprises administering to the subject an
effective amount of a Compound of Formula (I) or a pharmaceutically
acceptable salt thereof.
[0103] (cc) A method of treating HIV infection and/or reducing the
likelihood or severity of symptoms of HIV infection in a subject in
need thereof which comprises administering to the subject an
effective amount of a Compound of Formula (I) or a pharmaceutically
acceptable salt thereof.
[0104] (dd) The method of (cc), wherein the Compound of Formula (I)
or pharmaceutically acceptable salt thereof, is administered in
combination with an effective amount of at least one second
therapeutic agent selected from the group consisting of HIV
antiviral agents, immunomodulators, and anti-infective agents.
[0105] (ee) The method of (dd), wherein the HIV antiviral agent is
an antiviral selected from the group consisting of HIV protease
inhibitors and HIV NNRTI inhibitors.
[0106] (ff) A method of inhibiting HIV replication in a subject in
need thereof which comprises administering to the subject the
pharmaceutical composition of (w) (x) or (y) or the combination of
(z) or (aa).
[0107] (gg) A method of treating HIV infection and/or reducing the
likelihood or severity of symptoms of HIV infection in a subject in
need thereof which comprises administering to the subject the
pharmaceutical composition of (w) (x) or (y) or the combination of
(z) or (aa).
[0108] The present invention also includes a compound of the
present invention for use (i) in, (ii) as a medicament for, or
(iii) in the preparation of a medicament for: (a) medicine; (b)
inhibiting HIV replication or (c) treating HIV infection and/or
reducing the likelihood or severity of symptoms of HIV infection.
In these uses, the compounds of the present invention can
optionally be employed in combination with one or more second
therapeutic agents selected from HIV antiviral agents,
anti-infective agents, and immunomodulators.
[0109] Additional embodiments of the invention include the
pharmaceutical compositions, combinations and methods set forth in
(a)-(gg) above and the uses set forth in the preceding paragraph,
wherein the compound of the present invention employed therein is a
compound of one of the embodiments, aspects, classes, sub-classes,
or features of the compounds described above. In all of these
embodiments, the compound may optionally be used in the form of a
pharmaceutically acceptable salt or hydrate as appropriate.
[0110] It is further to be understood that the embodiments of
compositions and methods provided as (a) through (gg) above are
understood to include all embodiments of the compounds, including
such embodiments as result from combinations of embodiments.
[0111] Non-limiting examples of the Compounds of Formula (I)
include compounds 1A-18B as set forth in the Examples below, and
pharmaceutically acceptable salts thereof.
[0112] Methods for Making the Compounds of Formula (I)
[0113] The Compounds of Formula (I) may be prepared from known or
readily prepared starting materials, following methods known to one
skilled in the art of organic synthesis. Methods useful for making
the Compounds of Formula (I) are set forth in the Examples below
and generalized in the Schemes below. Alternative synthetic
pathways and analogous structures will be apparent to those skilled
in the art of organic synthesis.
General List of Abbreviations
[0114] Abbreviations and acronyms employed herein include the
following:
TABLE-US-00001 Ac Acetyl aq Aqueous AUC Area under the curve BAST
Bis(2-methoxyethyl)aminosulfur trifluoride Bu Butyl Bz Benzoyl
DBDMH 1,3-Dibromo-5,5-dimethylhydantoin DCM dichloromethane DHP
3,4-dihydro-2H-pyran DIEA, DIPEA N,N-diisopropylethylamine or
Hunig's base DMF dimethylformamide DMP Dess-Martin periodinane dppf
1,1'-Bis(diphenylphosphino)ferrocene DMSO dimethyl sulfoxide EDCI
N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride Et
Ethyl EtOH Ethanol EtOAc ethyl acetate g Grams GI gastrointenstinal
h Hour HIV human immunodeficiency virus HPBCD hvdroxypropyl
.beta.-cyclodextrin HPLC high-performance liquid chromatography
HOBT mCPBA Hz hertz IPA isopropanol IV intravenous iPr isopropyl L
liter LC liquid chromatography LC/MS liquid chromatography mass
spectrometry Me methyl MeOH methanol mg milligrams MHz megahertz
min minute .mu.L microliters mL milliliters mmol millimoles MS mass
spectrometry NBS N-Bromosuccinimide NHS normal human serum NIS
N-Iodosuccinimide NMR nuclear magnetic resonance spectroscopy PBMC
peripheral blood mononuclear cell Ph phenyl P.O. oral PTSA
para-toluenesulfonic acid Pr propyl RT or rt room temperature
(ambient, about 25.degree. C.) sat or sat'd saturated SFC
supercritical fluid chromatography TBAF Tetra-n-butylammonium
fluoride TBDPSCl tert-Butyldiphenylchlorosilane tBu tert-butyl TEA
triethylamine (Et.sub.3N) TEMED tetramethylethylenediamine TFA
trifluoroacetic acid TFV Tenofovir TFV-MP Tenofovir monophosphoate
TFV-DP Tenofovir diphosphate THF tetrahydrofuran TMS
tetramethylsilane UPLC ultrahigh pressure liquid chromatography UV
ultraviolet UV/VIS ultraviolet/visible
General Procedures
[0115] Starting materials and intermediates are purchased or are
made using known procedures, or as otherwise illustrated. The
general route applied to the synthesis of compounds of Formula I is
described in the Schemes that follows. In some cases the order of
carrying out the reaction steps in the schemes may be varied to
facilitate the reaction or to avoid unwanted reaction products.
[0116] Reactions sensitive to moisture or air were performed under
nitrogen or argon using anhydrous solvents and reagents. The
progress of reactions was determined by either analytical thin
layer chromatography (TLC) usually performed with E. Merck
pre-coated TLC plates, silica gel 60E-254, layer thickness 0.25 mm
or liquid chromatography-mass spectrometry (LC/MS).
[0117] Typically the analytical LC-MS system used consisted of a
Waters ZQ.TM. platform with electrospray ionization in positive ion
detection mode with an Agilent 1100 series HPLC with autosampler.
The column was commonly a Waters Xterra MS C18, 3.0.times.50 mm, 5
.mu.m or a Waters Acquity UPLC.RTM. BEH C18 1.0.times.50 mm, 1.7
.mu.m. The flow rate was 1 mL/min, and the injection volume was 10
.mu.L. UV detection was in the range 210-400 nm. The mobile phase
consisted of solvent A (water plus 0.05% TFA) and solvent B (MeCN
plus 0.05% TFA) with a gradient of 100% solvent A for 0.7 min
changing to 100% solvent B over 3.75 min, maintained for 1.1 min,
then reverting to 100% solvent A over 0.2 min.
[0118] Preparative HPLC purifications were usually performed using
either a mass spectrometry directed system or a non-mass guided
system. Usually they were performed on a Waters Chromatography
Workstation configured with LC-MS System consisting of: Waters
ZQ.TM. single quad MS system with Electrospray Ionization, Waters
2525 Gradient Pump, Waters 2767 Injecto/Collector, Waters 996 PDA
Detector, the MS Conditions of: 150-750 amu, Positive Electrospray,
Collection Triggered by MS, and a Waters SUNFIRE.RTM. C-18 5
micron, 30 mm (id).times.100 mm column. The mobile phases consisted
of mixtures of acetonitrile (10-100%) in water containing 0.1% TFA.
Flow rates were maintained at 50 mL/min, the injection volume was
1800 .mu.L, and the UV detection range was 210-400 nm. An alternate
preparative HPLC system used was a Gilson Workstation consisting
of: Gilson GX-281 Injector/Collector, Gilson UV/VIS-155 Detector,
Gilson 333 and 334 Pumps, and either a Phenomenex Gemini-NX C-18 5
micron, 50 mm (id).times.250 mm column or a Waters XBridge.TM. C-18
5 micron OBD.TM., 30 mm (id).times.250 mm column. The mobile phases
consisted of mixtures of acetonitrile (0-85%) in water containing
0.1% TFA. Flow rates were maintained at 50 mL/min for the Waters
Xbridge.TM. column and 90 mL/min for the Phenomenex Gemini column.
The injection volume ranged from 1000-8000 .mu.L, and the UV
detection range was 210-400 nm. Mobile phase gradients were
optimized for the individual compounds. Reactions performed using
microwave irradiation were normally carried out using an Emrys
Optimizer manufactured by Personal Chemistry, or an Initiator
manufactured by Biotage. Concentration of solutions was carried out
on a rotary evaporator under reduced pressure. Flash chromatography
was usually performed using either a Biotage.RTM. Flash
Chromatography apparatus (Dyax Corp.), an ISCO CombiFlash.RTM. Rf
apparatus, or an ISCO CombiFlash.RTM. Companion XL on silica gel
(32-63 microns, 60 .ANG. pore size) in pre-packed cartridges of the
size noted. .sup.1H NMR spectra were acquired at 500 MHz
spectrometers in CDCl.sub.3 solutions unless otherwise noted.
Chemical shifts were reported in parts per million (ppm).
Tetramethylsilane (TMS) was used as internal reference in
CD.sub.3Cl solutions, and residual CH.sub.3OH peak or TMS was used
as internal reference in CD.sub.3OD solutions. Coupling constants
(J) were reported in hertz (Hz). Chiral analytical chromatography
was most commonly performed on one of CHIRALPAK.RTM. AS,
CHIRALPAK.RTM.AD, CHIRALCEL.RTM. OD, CHIRALCEL.RTM. IA, or
CHIRALCEL.RTM. OJ columns (250.times.4.6 mm) (Daicel Chemical
Industries, Ltd.) with noted percentage of either ethanol in hexane
(% Et/Hex) or isopropanol in heptane (% IPA/Hep) as isocratic
solvent systems. Chiral preparative chromatography was conducted on
one of CHIRALPAK AS, of CHIRALPAK AD, CHIRALCEL.RTM. OD,
CHIRALCEL.RTM.IA, CHIRALCEL.RTM. OJ columns (20.times.250 mm)
(Daicel Chemical Industries, Ltd.) with desired isocratic solvent
systems identified on chiral analytical chromatography or by
supercritical fluid (SFC) conditions.
[0119] Several methods for preparing the compounds of this
invention are also described in the Examples. Starting materials
and intermediates were purchased commercially from common catalog
sources or were made using known procedures, or as otherwise
illustrated.
Preparation of Intermediate A
##STR00006## ##STR00007##
[0120] Step 1
3-(benzyloxy)-4-oxo-1,4-dihydropyridine-2-carboxylic acid
##STR00008##
[0122] 3-(benzyloxy)-4-oxo-4H-pyran-2-carboxylic acid (20 g, 81.23
mmol) was dissolved in NH.sub.3.H.sub.2O (150 mL, 28% aqueous,
81.23 mmol), and the reaction was stirred at 25.degree. C. for 20
h. The resulting mixture was concentrated in vacuo, and to the
resulting residue was added 80 mL aqueous 2 M HCl. The suspension
was filtered through a Celite pad, and the filter cake was
collected and dried under high vacuum to provide the title
compound, which is used to the next step without further
purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=7.71 (d,
J=6.4 Hz, 1H), 7.49-7.43 (m, 2H), 7.38-7.28 (m, 3H), 6.54 (d, J=5.7
Hz, 1H), 5.09 (s, 2H). LRMS (M+H).sup.+: 246.0.
Step 2
3-(benzyloxy)-4-oxo-1,4-dihydropyridine-2-carboxamide
##STR00009##
[0124] To a solution of
3-(benzyloxy)-4-oxo-1,4-dihydropyridine-2-carboxylic acid (48 g,
195.73 mmol) in DMF (400 mL) was added DIEA (180 mL, 1.02 mol),
HOBT (41.33 g, 305.83 mmol), and EDC (78.17 g, 407.78 mmol) at
0.degree. C. The reaction mixture was stirred at 25.degree. C. for
1 h, then solid NH.sub.4Cl (54.53 g, 1.02 mol) was added in
portions, and the reaction mixture was stirred at 25.degree. C. for
3 h. The resulting mixture was filtered and the filtrate was
concentrated in vacuo. The resulting residue was purified by
preparative HPLC (Column: Phenomenex Synergi Max-RP 150*50 mm*10
.mu.m; Conditions: water (0.1% TFA)/MeCN; begin B 0%, end B 30%;
gradient time 20 min; 100% B hold time 5 min, flowrate 120 mL/min)
to provide the title compound. .sup.1H NMR (400 MHz, CD.sub.3OD):
.delta.=7.91 (d, J=6.6 Hz, 1H), 7.47-7.41 (m, 2H), 7.35 (m, 3H),
6.88 (d, J=6.6 Hz, 1H), 5.47 (s, 2H). LRMS (M+H).sup.+:
245.0..sup.+
Step 3
1-amino-3-(benzyloxy)-4-oxo-1,4-dihydropyridine-2-carboxamide
##STR00010##
[0126] A mixture of
3-(benzyloxy)-4-oxo-1,4-dihydropyridine-2-carboxamide (1.5 g, 6.14
mmol) and K.sub.2CO.sub.3 (3.39 g, 24.56 mmol) in MeCN (30 mL) was
cooled to 0.degree. C. O-(2,4-dinitrophenyl)-hydroxylamine (2.45 g,
12.28 mmol) was then added as a solid in portions. The mixture was
stirred at 28.degree. C. for 16 h, and the resulting mixture was
concentrated in vacuo directly. The resulting crude product was
purified by gradient elutions on SiO.sub.2 (ISCO, 40-g RediSep Gold
column, 0 to 20% MeOH/DCM over 1 h, dry-loaded) to provide the
title compound. LRMS (M+H).sup.+: 260.0.
Step 4
5-(benzyloxy)-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione
(Intermediate A)
##STR00011##
[0128] To a solution of
1-amino-3-(benzyloxy)-4-oxo-1,4-dihydropyridine-2-carboxamide (2.0
g, 7.71 mmol) in DMF (30 mL) and AcOH (30 mL) was added
paraformaldehyde (198 mg, .about.0.9 eq). The mixture was stirred
at 100.degree. C. for 2 h, and then the resulting mixture was
concentrated in vacuo. The resulting crude product was slurried
with CH.sub.2Cl.sub.2 (20 mL), and the resulting suspension was
filtered through a celite pad. The filter cake was collected and
dried in vacuo to provide INTERMEDIATE A. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta.=8.96 (s, 1H), 7.73 (d, J=7.7 Hz, 1H),
7.60-7.50 (m, 2H), 7.40-7.26 (m, 4H), 6.21 (d, J=7.7 Hz, 1H), 5.09
(s, 2H), 4.38 (dd, J=3.6, 7.6 Hz, 2H). LRMS (M+H).sup.+: 272.1.
Preparation of Intermediate B
##STR00012##
[0129] Step 1
Dimethyl 3-((tert-butyldiphenylsilyl)oxy)pentanedioate
##STR00013##
[0131] To a solution of dimethyl 3-hydroxypentanedioate (20 g, 114
mmol) in CH.sub.2Cl.sub.2 (100 mL) was added 1H-imidazole (15.46 g,
227 mmol) and TBDPSCl (46.8 g, 170 mmol) at 25.degree. C. The
solution was stirred at 25.degree. C. for 3 h, and the mixture was
filtrated and the filter cake was washed by CH.sub.2Cl.sub.2 (100
mL). The filtrate was concentrated in vacuo to afford the crude
product, which was purified by gradient elution on SiO.sub.2 (ISCO,
120 g RediSep Gold column, 0 to 10% EtOAc/Pet.ether over 90 min,
dry-loaded) to afford the title compound. LRMS (M+H).sup.+:
437.2.
Step 2
3-((tert-buty ldiphenylsilyl)oxy)pentane-1,5-diol
##STR00014##
[0133] To a solution of dimethyl
3-((tert-butyldiphenylsilyl)oxy)pentanedioate (30.166 g, 72.8 mmol)
in THF (125 mL) was added LiBH.sub.4 (6.34 g, 291 mmol) at
0.degree. C. The solution was stirred at 25.degree. C. for 2 h, and
then the reaction was quenched with 1M aq. HCl (20 mL) at 0.degree.
C. The mixture was extracted with ethyl acetate (3.times.50 mL),
the combined organic layers were dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The crude product was purified
by gradient elution on SiO.sub.2 (ISCO, 330 g RediSep Gold column,
0 to 50% EtOAc/Pet.ether over 90 min, dry-loaded) to afford the
title compound. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta.=7.64-7.72 (m, 4H), 7.33-7.46 (m, 6H), 4.07-4.12 (m, 1H),
3.52-3.65 (m, 4H), 1.64-1.87 (m, 4H), 1.05 (s, 9H). LRMS
(M+H).sup.+: 359.0
Step 3
Tert-butyl((1,5-dibromopentan-3-yl)oxy)diphenylsilane
##STR00015##
[0135] To a solution of
3-((tert-butyldiphenylsilyl)oxy)pentane-1,5-diol (5.319 g, 14.83
mmol) in CH.sub.2Cl.sub.2 (100 mL) was added CBr.sub.4 (14.76 g,
44.5 mmol) and PPh.sub.3 (11.67 g, 44.5 mmol) at 0.degree. C., the
mixture was stirred at 25.degree. C. for 6 h. The resulting mixture
was quenched with saturated aq. Na.sub.2SO.sub.3 (30 mL), and the
resulting mixture was extracted with EtOAc (3.times.100 mL), washed
with brine (20 mL), dried over Na.sub.2SO.sub.4 and filtered. The
crude product was purified by gradient elution on SiO.sub.2 (ISCO,
330 g silica Flash Column, 0 to 10% EtOAc/Pet.ether over 90 min,
dry-loaded) to provide the title compound. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta.=7.69 (dd, J=7.9, 1.5 Hz, 4H), 7.37-7.48 (m,
6H), 4.02 (d, J=5.7 Hz, 1H), 3.33 (t, J=7.2 Hz, 4H), 1.94-2.10 (m,
4H), 1.07 (s, 9H).
PREPARATION OF EXAMPLES
Example 1
##STR00016##
[0136] Step 1
9-(benzyloxy)-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazec-
ine-8,10-dione
##STR00017##
[0138] A mixture of INTERMEDIATE A (1.1 g, 4.05 mmol) and
1,5-dibromopentane (979.02 mg, 4.26 mmol) in DMF (100 mL) was
cooled to 0.degree. C. Powdered KOH (1.82 g, 32.44 mmol) was
suspended in DMF (50 mL) and added dropwise (via syringe) to the
stirred mixture at 0.degree. C. over a period of 2 h. At this time,
the resulting mixture was diluted with water (50 mL), extracted
with CH.sub.2Cl.sub.2 (3.times.60 mL), washed with brine (20 mL),
dried over Na.sub.2SO.sub.4 and filtered. The filtrate was
concentrated in vacuo. The residue was purified by gradient elution
on SiO.sub.2 (ISCO, 40-g RediSep Gold column, 0 to 10% MeOH/EtOAc
over 30 min, dry-loaded) to provide the title compound. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta.=7.54 (d, J=7.7 Hz, 2H), 7.32-7.19 (m,
4H), 6.36-6.31 (m, 1H), 5.45 (dd, J=2.4, 10.4 Hz, 1H), 5.12 (dd,
J=1.3, 10.1 Hz, 1H), 4.58-4.49 (m, 1H), 4.45-4.36 (m, 1H),
4.33-4.22 (m, 1H), 3.32 (dd, J=9.8, 11.8 Hz, 1H), 3.08-2.96 (m,
1H), 2.82-2.73 (m, 1H), 2.05-1.95 (m, 1H), 1.74-1.63 (m, 4H), 1.55
(m, 1H). LRMS (M+H).sup.+: 340.1.
Step 2
9-(benzyloxy)-11-bromo-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[1,2-b][1,2,-
5]triazecine-8,10-dione
##STR00018##
[0140] A mixture of
9-(benzyloxy)-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triaze-
cine-8,10-dione (600 mg, 1.77 mmol) in CH.sub.2Cl.sub.2 (6 mL) was
cooled to 0.degree. C., then NBS (943.95 mg, 5.3 mmol) was added in
portions as a solid. The mixture was stirred at 0.degree. C. for 45
min, and the resulting mixture was quenched with saturated aq
Na.sub.2SO.sub.3 (8 mL), extracted with CH.sub.2Cl.sub.2
(3.times.10 mL), washed with brine (10 mL), dried over
Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated in
vacuo. The residue was purified by normal phase chromatography
(ISCO, 20-g RediSep Gold column, 0 to 10% MeOH/EtOAc over 2 h,
dry-loaded) to provide the title compound. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta.=7.85 (s, 1H), 7.65 (d, J=6.6 Hz, 2H), 7.38-7.29
(m, 3H), 5.55 (d, J=10.1 Hz, 1H), 5.19 (d, J=10.1 Hz, 1H),
4.72-4.63 (m, 1H), 4.50 (d, J=14.5 Hz, 1H), 4.41-4.32 (m, 1H),
3.51-3.38 (m, 1H), 3.21-3.11 (m, 1H), 2.90-2.81 (m, 1H), 2.13-2.03
(m, 1H), 1.79 (br d, J=11.0 Hz, 4H), 1.64-1.59 (m, 1H). LRMS
(M+H).sup.+: 418.1.
Step 3
9-(benzyloxy)-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexahydro--
2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00019##
[0142] To a solution of
9-(benzyloxy)-11-bromo-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[1,2-b][1,2-
,5]triazecine-8,10-dione (591 mg, 1.41 mmol) in DMSO (6 mL) was
added (2,4,6-trifluorophenyl)methanamine (341.48 mg, 2.12 mmol),
Pd(PPh.sub.3).sub.4 (489.82 mg, 423.87 .mu.mol) and DIEA (1.72 mL,
9.89 mmol). The mixture was stirred at 90.degree. C. for 1.5 h
under CO (15 psi). The mixture was filtered and purified by
preparative HPLC (Column: Phenomenex Synergi Max-RP 150*50 mm, 10
.mu.m; Conditions: water (0.1% TFA)/MeCN; begin B 0%, end B 30%;
gradient time 20 min; 100% B hold time 5 min, flowrate 120 mL/min)
to provide the title compound. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=10.31 (t, J=5.4 Hz, 1H), 8.46 (s, 1H), 7.54-7.48 (m, 2H),
7.31-7.20 (m, 3H), 6.67-6.54 (m, 2H), 5.42 (d, J=10.1 Hz, 1H), 5.13
(d, J=9.9 Hz, 1H), 4.64-4.44 (m, 4H), 4.30 (br dd, J=7.6, 14.0 Hz,
1H), 3.47-3.33 (m, 1H), 3.1-3.03 (m, 1H), 2.84-2.75 (m, 1H), 2.00
(dd, J=6.2, 10.6 Hz, 1H), 1.70 (m, 3H), 1.60 (m, 2H). LRMS
(M+H).sup.+: 527.2.
Step 4
9-(benzyloxy)-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexahydro--
2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00020##
[0144] Enantiomers were separated by preparative chiral SFC
(Column: YMC CHIRAL Amylose-C 250*30 mm, 5 .mu.m; Conditions:
CO.sub.2/IPA (0.1% NH.sub.4OH); begin B 40%, end B 40% (isocratic);
flowrate 60 mL/min; time: 16.2 min; peak 1 rt=9.25 min, peak 2
rt=12.5 min) to provide
9-(benzyloxy)-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexahydro-
-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide (PEAK
1, >99.9% ee) and
9-(benzyloxy)-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexahydro-
-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide (PEAK
2, 99.6% ee). LRMS (M+H).sup.+: 527.2.
Step 5
9-hydroxy-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexahydro-2H-1-
,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00021##
[0146] A solution of PEAK 1 of
9-(benzyloxy)-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexahydro-
-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide (14
mg, 0.027 mmol) from the preceding chiral separation and MgBr.sub.2
(14.69 mg, 0.080 mmol) in MeCN (1 mL) was stirred at 40.degree. C.
for 0.5 h. The mixture was purified directly by preparative HPLC
(Column: Boston Green ODS 150*30 mm, 5 .mu.m; Conditions: water
(0.1% TFA)/MeCN; begin B 23%, end B 53%; gradient time 8 min,
flowrate 30 mL/min) to provide 1A .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=10.33 (s, 1H), 8.48 (s, 1H), 6.68-6.64 (m, 2H), 4.86 (d,
J=14.4 Hz, 1H), 4.69-4.65 (m, 3H), 4.37-4.33 (m, 1H), 3.55-3.51 (m,
1H), 3.18-3.16 (m, 1H), 3.04-3.00 (m, 1H), 2.07 (s, 1H), 1.88-1.68
(m, 5H). LRMS (M+H).sup.+: 437.1. PEAK 2 of
9-(benzyloxy)-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexahydro-
-2H-1,7-methano-pyrido[1,2-b][1,2,5]triazecine-11-carboxamide from
the preceding chiral separation was subjected to the same reaction
conditions to afford 1B. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=10.33 (s, 1H), 8.48 (s, 1H), 6.68-6.64 (m, 2H), 4.87 (d,
J=14.4 Hz, 1H), 4.69-4.63 (m, 3H), 4.36-4.34 (m, 1H), 3.52-3.50 (m,
1H), 3.18-3.16 (m, 1H), 3.04-3.00 (m, 1H), 2.06 (s, 1H), 1.78-1.66
(m, 5H). LRMS (M+H).sup.+: 437.1.
Example 2
##STR00022## ##STR00023##
[0147] Step 1
8-(benzyloxy)-2,3,4,5-tetrahydro-1,6-methanopyrido[1,2-b][1,2,5]triazonine-
-7,9-dione
##STR00024##
[0149] Solid KOH (0.776 g, 13.82 mmol) was suspended in DMSO (40
mL), and the solution was stirred at 70.degree. C. until most solid
particles disappeared. Then the cooled base solution was added
dropwise into a solution of INTERMEDIATE A (1.271 g, 4.69 mmol) and
1,4-dibromobutane (1.01 g, 4.69 mmol) in DMSO (150 mL) over 2.5 h
at 20.degree. C. The reaction was stirred at 20.degree. C. for
another 1 h. The mixture was poured into water (200 mL) and
extracted with MeOH/DCM (1:10, 3.times.200 mL). The combined
organic layers were washed with brine, dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The residue was purified by
elution on SiO.sub.2 (isocratic, 5% MeOH in CH.sub.2Cl.sub.2) to
provide the title compound. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=7.87 (s, 1H), 7.43 (d, J=7.6 Hz, 2H), 7.36-7.32 (m, 4H),
5.32 (d, J=9.6 Hz, 1H), 5.27 (d, J=9.6 Hz, 1H), 4.63 (s, 1H), 4.47
(s, 1H), 4.30 (s, 1H), 3.39-3.47 (m, 2H), 3.13 (s, 1H), 2.24-2.21
(m, 1H), 1.87-1.56 (m, 3H). LRMS (M+H).sup.+: 340.0.
Step 2
8-(benzyloxy)-10-bromo-2,3,4,5-tetrahydro-1,6-methanopyrido[1,2-b][1,2,5]t-
riazonine-7,9-dione
##STR00025##
[0151] To a stirred yellow solution of
8-(benzyloxy)-2,3,4,5-tetrahydro-1,6-methanopyrido[1,2-b][1,2,5]triazonin-
e-7,9-dione (40 mg, 0.123 mmol) in CH.sub.2Cl.sub.2 (1 mL) was
added NBS (43.8 mg, 0.246 mmol) at 0.degree. C. The mixture was
stirred at 20.degree. C. for 0.5 h and then purified directly by
preparative TLC eluting with 5% MeOH in CH.sub.2Cl.sub.2 to provide
the title compound. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.=7.89
(s, 1H), 7.14 (d, J=7.6 Hz, 2H), 7.36-7.29 (m, 3H), 5.61 (d, J=10.0
Hz, 1H), 5.16 (d, J=10.0 Hz, 1H), 4.55 (d, J=10.0 Hz, 1H),
4.43-4.21 (m, 1H), 4.19 (d, J=10.0 Hz, 1H), 3.37-3.30 (m, 2H),
3.27-3.26 (m, 1H), 2.18-2.16 (m, 1H), 1.81-1.58 (m, 3H). LRMS
(M+H).sup.+: 406.1.
Step 3
8-(benzyloxy)-N-(2,4-difluorobenzyl)-7,9-dioxo-2,3,4,5,7,9-hexahydro-1,6-m-
ethanopyrido[1,2-b][1,2,5]triazonine-10-carboxamide
##STR00026##
[0153] A mixture of
8-(benzyloxy)-10-bromo-2,3,4,5-tetrahydro-1,6-methanopyrido[1,2-b][1,2,5]-
triazonine-7,9-dione (52 mg, 0.129 mmol),
(2,4-difluorophenyl)methanamine (33.1 mg, 0.232 mmol),
Pd(PPh.sub.3).sub.4 (74.3 mg, 0.064 mmol) and DIEA (0.067 mL, 0.386
mmol) in DMSO (1 mL) was stirred under CO balloon (15 psi) at
90.degree. C. for 2 h. The mixture was filtered and the filtrate
purified by preparative HPLC (Column: YMC-Actus Pro C18 150*30 mm,
5 .mu.m; Conditions: water (0.1% TFA)/MeCN; begin B 0%, end B 30%;
gradient time 11 min; 100% B hold time 1.1 min, flowrate 40 mL/min)
to provide the title compound. LRMS (M+H).sup.+: 495.2.
Step 4
8-(benzyloxy)-N-(2,4-difluorobenzyl)-7,9-dioxo-2,3,4,5,7,9-hexahydro-1,6-m-
ethanopyrido[1,2-b][1,2,5]triazonine-10-carboxamide
##STR00027##
[0155] Enantiomers were separated by preparative chiral SFC
(Column: CHIRALCEL-AS 250*30 mm, 5 .mu.m; Conditions: isocratic
CO.sub.2/EtOH (0.1% NH.sub.4OH); begin B 45%, end B 45%
(isocratic); flowrate 50 mL/min; peak 1 rt=0.8 min, peak 2 rt=1.2
min) to provide
8-(benzyloxy)-N-(2,4-difluorobenzyl)-7,9-dioxo-2,3,4,5,7,9-hexahydro-1,6--
methanopyrido[1,2-b][1,2,5]triazonine-10-carboxamide (PEAK 1,
>99.9% ee) and
8-(benzyloxy)-N-(2,4-difluorobenzyl)-7,9-dioxo-2,3,4,5,7,9-hexahy-
dro-1,6-methanopyrido[1,2-b][1,2,5]triazonine-10-carboxamide (PEAK
2, >99.9% ee). LRMS (M+H).sup.+: 495.2.
Step 5
N-(2,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9-hexahydro-1,6-metha-
nopyrido[1,2-1)][1,2,5]triazonine-10-carboxamide
##STR00028##
[0157] A solution of PEAK 1 of
8-(benzyloxy)-N-(2,4-difluorobenzyl)-7,9-dioxo-2,3,4,5,7,9-hexahydro-1,6--
methanopyrido[1,2-b][1,2,5]triazonine-10-carboxamide (10 mg, 0.020
mmol) and MgBr.sub.2 (11.17 mg, 0.061 mmol) in MeCN (1 mL) was
stirred at 20.degree. C. for 2 h. The mixture was purified directly
by preparative HPLC (Column: DuraShell 150*30 mm, 5 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 28%, end B 58%; gradient
time 10 min, 100% B hold time 2.5 min, flowrate 30 mL/min) to
provide 2A. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.=10.34 (s,
1H), 8.57 (s, 1H), 7.40-7.34 (m, 1H), 6.85-6.70 (m, 2H), 4.85 (d,
J=14 Hz, 1H), 4.46 (s, 2H), 4.43-4.37 (m, 2H), 3.41-4.39 (m, 2H),
3.31-3.23 (m, 1H), 2.31-2.25 (m, 1H), 1.76-1.75 (m, 1H), 1.74-1.58
(m, 2H). LRMS (M+H).sup.+: 405.1. PEAK 2 of
8-(benzyloxy)-N-(2,4-difluorobenzyl)-7,9-dioxo-2,3,4,5,7,9-hexahydro-1,6--
methanopyrido[1,2-b][1,2,5]triazonine-10-carboxamide from the
preceding chiral separation was subjected to the same reaction
conditions to afford 2B. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=10.34 (s, 1H), 8.57 (s, 1H), 7.40-7.34 (m, 1H), 6.85-6.79
(m, 2H), 4.82 (d, J=14 Hz, 1H), 4.66 (s, 2H), 4.43-4.37 (m, 2H),
3.41-3.39 (m, 2H), 3.31-3.23 (m, 1H), 2.31-2.25 (m, 1H), 1.76-1.75
(m, 1H), 1.74-1.60 (m, 2H). LRMS (M+H).sup.+: 405.1.
Example 3
##STR00029## ##STR00030##
[0158] Step 1
5-(benzyloxy)-1-(6-bromohexyl)-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-
-4,6-dione
##STR00031##
[0160] Solid KOH (0.371 g, 6.61 mmol) was suspended in DMSO (20
mL), and the solution was stirred at 70.degree. C. until most solid
particles disappeared. INTERMEDIATE A (1.4 g, 5.16 mmol) and
1,6-dibromohexane (1.89 g, 7.74 mmol) were added, and the reaction
mixture was stirred at 20.degree. C. for 8 h. The mixture was
poured into water (5 mL) and extracted with CH.sub.2Cl.sub.2
(3.times.5 mL). The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered, concentrated in vacuo and the crude
product was purified by gradient elution on SiO.sub.2 (isocratic,
0% to 10% MeOH in 3:1 EtOAc/EtOH) to provide the title compound.
LRMS (M+H).sup.+: 434.0, 436.0.
Step 2
10-(benzyloxy)-2,3,4,5,6,7-hexahydro-1,8-methanopyrido[1,2-b][1,2,5]triaza-
cycloundecine-9,11-dione
##STR00032##
[0162] To a solution of
5-(benzyloxy)-1-(6-bromohexyl)-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazin-
e-4,6-dione (1.1 g, 2.53 mmol) in DMSO (20 mL) was added powdered
KOH (0.213 g, 3.80 mmol) in one portion at 20.degree. C. and
stirred at this temperature for 4 h. The mixture was poured into
water (10 mL) and extracted with CH.sub.2Cl.sub.2 (3.times.10 mL).
The combined organic layers were purified directly by preparative
HPLC (Phenomenex Luna C18 250*80 mm, 10 .mu.m; Conditions: water
(0.1% TFA)/MeCN; begin B 15%, end B 35%; flowrate 250 mL/min) to
provide the title compound. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=7.95 (br d, J=7.3 Hz, 1H), 7.59 (br d, J=7.3 Hz, 1H),
7.49-7.56 (m, 2H), 7.33-7.41 (m, 3H), 5.40 (d, J=9.9 Hz, 1H), 5.29
(d, J=9.9 Hz, 1H) 4.72 (s, 2H), 4.30-4.47 (m, 1H), 3.23-3.46 (m,
2H), 2.76-2.86 (m, 1H), 1.48-2.11 (m, 8H). LRMS (M+H).sup.+:
354.2.
Step 3
10-(benzyloxy)-12-bromo-2,3,4,5,6,7-hexahydro-1,8-methanopyrido[1,2-b][1,2-
,5]triazacycloundecine-9,11-dione
##STR00033##
[0164] To a stirred solution of
10-(benzyloxy)-2,3,4,5,6,7-hexahydro-1,8-methanopyrido[1,2-b][1,2,5]triaz-
acycloundecine-9,11-dione (350 mg, 0.990 mmol) in CH.sub.2Cl.sub.2
(5 mL) was added 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione
(170 mg, 0.594 mmol) in one portion at 20.degree. C., and the
mixture was stirred at 20.degree. C. for 0.5 h. The reaction
mixture was purified directly by preparative TLC eluting with EtOAc
to provide the title compound. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=7.86 (s, 1H), 7.65 (d, J=7.1 Hz, 2H), 7.28-7.38 (m, 3H),
5.51 (d, J=9.9 Hz, 1H), 5.19 (d, J=9.9 Hz, 1H), 4.58-4.74 (m, 2H),
3.16-3.35 (m, 2H), 2.67-2.77 (m, 2H), 1.48-1.99 (m, 8H). LRMS
(M+H).sup.+: 432.0, 434.0.
Step 4
10-(benzyloxy)-N-(2,4-difluorobenzyl)-9,11-dioxo-2,3,4,5,6,7,9,11-octahydr-
o-1,8-methanopyrido[1,2-b][1,2,5]triazacycloundecine-12-carboxamide
##STR00034##
[0166] A mixture of
10-(benzyloxy)-12-bromo-2,3,4,5,6,7-hexahydro-1,8-methanopyrido[1,2-b][1,-
2,5]triazacycloundecine-9,11-dione (100 mg, 0.231 mmol),
(2,4-difluorophenyl)-methanamine (66.2 mg, 0.463 mmol),
Pd(Ph.sub.3P).sub.4 (134 mg, 0.116 mmol) and DIEA (0.121 mL, 0.694
mmol) in DMSO (0.5 mL) was stirred under CO balloon (15 psi) at
90.degree. C. for 5 h. The mixture was filtered and the filtrate
was diluted with water (5 mL) and extracted with EtOAc (3.times.10
mL). The combined organic layer was washed with brine (5 mL), dried
over anhydrous Na.sub.2SO.sub.4 and filtered. The organic layer was
concentrated in vacuo and purified by preparative TLC eluting with
EtOAc to provide the title compound. LRMS (M+H).sup.+: 523.2.
Step 5
10-(benzyloxy)-N-(2,4-difluorobenzyl)-9,11-dioxo-2,3,4,5,6,7,9,11-octahydr-
o-1,8-methanopyrido[1,2-b][1,2,5]triazacycloundecine-12-carboxamide
##STR00035##
[0168] Enantiomers were separated by preparative chiral SFC
(Column: CHIRALCEL-OD 250*30 mm, 10 .mu.m; Conditions: isocratic
CO.sub.2/EtOH (0.1% NH.sub.4OH); begin B 50%, end B 50%
(isocratic); flowrate 80 mL/min; peak 1 rt=0.74 min, peak 2 rt=1.26
min) to provide
10-(benzyloxy)-N-(2,4-difluorobenzyl)-9,11-dioxo-2,3,4,5,6,7,9,11-octahyd-
ro-1,8-methanopyrido[1,2-b][1,2,5]tria-zacycloundecine-12-carboxamide
(PEAK 1, >99.9% ee) and
10-(benzyloxy)-N-(2,4-difluorobenzyl)-9,11-dioxo-2,3,4,5,6,7,9,11-octahyd-
ro-1,8-methanopyrido[1,2-b][1,2,5]triaza-cycloundecine-12-carboxamide
(PEAK 2, 99.02% ee). LRMS (M+H).sup.+: 523.2.
Step 6
N-(2,4-difluorobenzyl)-10-hydroxy-9,11-dioxo-2,3,4,5,6,7,9,11-octahydro-1,-
8-methanopyrido[1,2-b][1,2,5]triazacycloundecine-12-carboxamide
##STR00036##
[0170] To a solution of PEAK 1 of
10-(benzyloxy)-N-(2,4-difluorobenzyl)-9,11-dioxo-2,3,4,5,6,7,9,11-octahyd-
ro-1,8-methanopyrido[1,2-b][1,2,5]triazacycloundecine-12-carboxamide
(40 mg, 0.077 mmol) in MeCN (2 mL) was added MgBr.sub.2 (14.09 mg,
0.077 mmol), and the mixture was stirred at 40.degree. C. for 2 h.
The mixture was filtered and purified directly by reverse phase
preparative HPLC (Column: Boston Green ODS 150*30 mm, 5 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 38%, end B 68%; gradient
time 10 min; 100% B hold time 2.5 min, flowrate 25 mL/min) to
provide 3A. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.=10.39 (br s,
1H), 8.46 (s, 1H), 7.30-7.41 (m, 1H), 6.71-6.86 (m, 2H), 4.72-4.91
(m, 2H), 4.55-4.67 (m, 2H), 4.32-4.44 (m, 1H), 3.19-3.32 (m, 2H),
2.77-2.88 (m, 1H), 1.75-1.92 (m, 3H), 1.46-1.66 (m, 5H). LRMS
(M+H).sup.+: 433.2. PEAK 2 of
10-(benzyloxy)-N-(2,4-difluorobenzyl)-9,11-dioxo-2,3,4,5,6,7,9,11-octahyd-
ro-1,8-methanopyrido[1,2-b][1,2,5]triazacycloundecine-12-carboxamide
from the preceding chiral separation was subjected to the same
reaction conditions to afford 3B. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=10.32-10.48 (m, 1H), 8.47 (s, 1H), 7.30-7.42 (m, 1H),
6.72-6.87 (m, 2H), 4.83-4.90 (m, 1H), 4.73-4.80 (m, 1H), 4.58-4.68
(m, 2H), 4.34-4.43 (m, 1H), 3.20-3.31 (m, 2H), 2.80-2.86 (m, 1H),
1.77-1.99 (m, 3H), 1.47-1.73 (m, 5H). LRMS (M+H).sup.+: 433.2.
Example 4
##STR00037##
[0171] Step 1
5-(benzyloxy)-1-(2-(2-bromoethoxy)ethyl)-2,3-dihydro-1H-pyrido[2,1-f][1,2,-
4]triazine-4,6-dione
##STR00038##
[0173] Solid KOH (124 mg, 2.21 mmol) was suspended in DMSO (25 mL),
and the solution was stirred at 70.degree. C. until most solid
particles disappeared. INTERMEDIATE A (300 mg, 1.11 mmol) and
1-bromo-2-(2-bromoethoxy)ethane (308 mg, 1.33 mmol) were added, and
the reaction mixture was stirred at 20.degree. C. for 3 h. The
mixture was poured into water (10 mL) and extracted with DCM
(3.times.10 mL). The combined organic layers were washed with
brine, dried over Na.sub.2SO.sub.4, filtered, the filtrate was
concentrated in vacuo. The crude residue was purified by
preparative TLC eluting with 10% MeOH in CH.sub.2Cl.sub.2 to
provide the title compound. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=7.79 (d, J=7.6 Hz, 1H), 7.51-7.46 (m, 2H), 7.36-7.32 (m,
3H), 7.15 (d, J=7.2 Hz, 1H), 5.24 (s, 2H), 4.62 (s, 1H), 3.80-3.78
(m, 2H), 3.68 (s, 4H), 3.51-3.49 (m, 2H). LRMS (M+H).sup.+:
424.1.
Step 2
9-(benzyloxy)-2,3,5,6-tetrahydro-1,7-methanopyrido[1,2-e][1,4,5,8]oxatriaz-
ecine-8,10-dione
##STR00039##
[0175] To a stirred yellow solution of
5-(benzyloxy)-1-(2-(2-bromoethoxy)ethyl)-2,3-dihydro-1H-pyrido[2,1-f][1,2-
,4]triazine-4,6-dione (40 mg, 0.095 mmol) in DMSO (1 mL) was added
KOH (42.5 mg, 0.76 mmol) in one portion at 20.degree. C., and then
the mixture was stirred at 20.degree. C. for 12 h. The mixture was
poured into water (10 mL) and extracted with DCM (3.times.10 mL).
The combined organic layers were dried over Na.sub.2SO.sub.4 and
filtered. The filtrate was concentrated under reduced pressure and
purified by preparative HPLC (Column: YMC-Actus Pro C18 150*30 mm,
5 .mu.m; Conditions: water (0.1% TFA)/MeCN; begin B 10%, end B 40%;
gradient time 11 min; 100% B hold time 1.1 min, flowrate 40 mL/min)
to provide the title compound. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=7.52-7.51 (m, 2H), 7.33 (d, J=7.6 Hz, 1H), 7.28-7.21 (m,
3H), 6.36 (d, J=7.6 Hz, 1H), 5.45 (d, J=10.4 Hz, 1H), 5.13 (d,
J=10.4 Hz, 1H), 4.78-4.67 (m, 2H), 4.12-4.00 (m, 1H), 3.96-3.92 (m,
2H), 3.84-3.82 (m, 1H), 3.56-3.55 (m, 1H), 3.41-3.40 (m, 1H),
3.24-3.23 (m, 1H), 2.98-2.97 (m, 1H). LRMS (M+H).sup.+: 342.2.
Step 3
9-(benzyloxy)-11-bromo-2,3,5,6-tetrahydro-1,7-methanopyrido[1,2-e][1,4,5,8-
]oxatriazecine-8,10-dione
##STR00040##
[0177] To a stirred solution of
9-(benzyloxy)-2,3,5,6-tetrahydro-1,7-methanopyrido[1,2-e][1,4,5,8]oxatria-
zecine-8,10-dione (25 mg, 0.073 mmol) in CH.sub.2Cl.sub.2 (1 mL)
was added NBS (39.1 mg, 0.22 mmol) in one portion at 0.degree. C.
The mixture was stirred at 20.degree. C. for 0.5 h. The mixture was
purified by preparative TLC eluting with 5% MeOH in
CH.sub.2Cl.sub.2 to provide the title compound. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta.=7.89 (s, 1H), 7.14 (d, J=7.6 Hz, 2H),
7.36-7.29 (m, 3H), 5.56 (d, J=10.4 Hz, 1H), 5.18 (d, J=10.4 Hz,
1H), 4.69-4.52 (m, 1H), 4.52-4.40 (m, 1H), 4.35 (d, J=14.5 Hz, 1H),
4.04-3.98 (m, 2H), 3.96-3.92 (m, 1H), 3.68-3.49 (m, 2H), 3.41-3.32
(m, 1H), 3.10-3.00 (m, 1H). LRMS (M+H).sup.+: 420.1.
Step 4
9-(benzyloxy)-N-(2,4-difluorobenzyl)-8,10-dioxo-2,3,5,6,8,10-hexahydro-1,7-
-methanopyrido[1,2-e][1,4,5,8]oxatriazecine-11-carboxamide
##STR00041##
[0179] A mixture of
9-(benzyloxy)-11-bromo-2,3,5,6-tetrahydro-1,7-methanopyrido[1,2-e][1,4,5,-
8]oxatriazecine-8,10-dione (20 mg, 0.048 mmol),
(2,4-difluorophenyl)methanamine (12.26 mg, 0.086 mmol),
Pd(PPh.sub.3).sub.4 (27.5 mg, 0.024 mmol) and DIEA (0.025 mL, 0.14
mmol) in DMSO (1 mL) was stirred under CO balloon (15 psi) at
90.degree. C. for 2 h. The mixture was filtered and the filtrate
was purified by preparative HPLC (Column: YMC-Actus Pro C18 150*30
mm, 5 .mu.m; Conditions: water (0.1% TFA)/MeCN; begin B 30%, end B
60%; gradient time 11 min; 100% B hold time 1.1 min, flowrate 40
mL/min) to provide the title compound. LRMS (M+H).sup.+: 511.0.
Step 5
9-(benzyloxy)-N-(2,4-difluorobenzyl)-8,10-dioxo-2,3,5,6,8,10-hexahydro-1,7-
-methanopyrido[1,2-e][1,4,5,8]oxatriazecine-11-carboxamide
##STR00042##
[0181] Enantiomers were separated by preparative chiral SFC
(Column: CHIRALCEL-AS 250*30 mm, 5 .mu.m; Conditions: isocratic
CO.sub.2/EtOH (0.1% NH.sub.4OH); begin B 40%, end B 40%
(isocratic); flowrate 50 mL/min; peak 1 rt=3.59 min, peak 2 rt=4.36
min) to provide
9-(benzyloxy)-N-(2,4-difluorobenzyl)-8,10-dioxo-2,3,5,6,8,10-hexahydro-1,-
7-methanopyrido[1,2-e][1,4,5,8]-oxa-triazecine-11-carboxamide (PEAK
1, >99.9% ee) and
9-(benzyloxy)-N-(2,4-difluorobenzyl)-8,10-dioxo-2,3,5,6,8,10-hexahydro-1,-
7-methanopyrido[1,2-e][1,4,5,8]oxa-triazecine-11-carboxamide (PEAK
2, 99.02% ee). LRMS (M+H).sup.+: 511.0.
Step 6
N-(2,4-difluorobenzyl)-9-hydroxy-8,10-dioxo-2,3,5,6,8,10-hexahydro-1,7-met-
hanopyrido[1,2-e][1,4,5,8]oxatriazecine-11-carboxamide
##STR00043##
[0183] To a solution of PEAK 1 of
9-(benzyloxy)-N-(2,4-difluorobenzyl)-8,10-dioxo-2,3,5,6,8,10-hexahydro-1,-
7-methanopyrido[1,2-e][1,4,5,8]oxatriazecine-11-carboxamide (8 mg,
0.016 mmol) in MeCN (1 mL) was added MgBr.sub.2 (14.43 mg, 0.078
mmol), and the mixture was stirred at 40.degree. C. for 2 h. The
mixture was filtered and purified directly by reverse phase
preparative HPLC (Column: Boston Green ODS 150*30 mm, 5 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 23%, end B 53%; gradient
time 8 min; flowrate 30 mL/min) to provide 4A. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta.=10.33 (s, 1H), 8.52 (s, 1H), 7.39-7.33 (m,
1H), 6.84-6.78 (m, 2H), 5.09 (d, J=13.6 Hz, 1H), 4.92 (d, J=13.6
Hz, 1H), 4.65-4.63 (m, 2H), 4.32-4.28 (m, 1H), 4.03-4.00 (m, 2H),
3.99-3.97 (m, 1H), 3.66-3.60 (m, 2H), 3.35-3.32 (m, 1H), 3.18-3.16
(m, 1H). LRMS (M+H).sup.+: 421.0. PEAK 2 of
9-(benzyloxy)-N-(2,4-difluorobenzyl)-8,10-dioxo-2,3,5,6,8,10-hexahydro-1,-
7-methanopyrido[1,2-e][1,4,5,8]-oxa-triazecine-11-carboxamide from
the preceding chiral separation was subjected to the same reaction
conditions to afford 4B. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.=10.32 (s, 1H), 8.51 (s, 1H), 7.38-7.32 (m, 1H), 6.83-6.77
(m, 2H), 5.08 (d, J=13.6 Hz, 1H), 4.91 (d, J=13.6 Hz, 1H),
4.64-4.63 (m, 2H), 4.31-4.27 (m, 1H), 4.02-3.99 (m, 2H), 3.98-3.97
(m, 1H), 3.65-3.59 (m, 2H), 3.35-3.31 (m, 1H), 3.17-3.15 (m, 1H).
LRMS (M+H).sup.+: 421.0.
Example 5
##STR00044##
[0184] Step 1
9-(benzyloxy)-4-((tert-butyldiphenylsilyl)oxy)-3,4,5,6-tetrahydro-2H-1,7-m-
ethanopyrido[1,2-b][1,2,5]triazecine-8,10-dione
##STR00045##
[0186] To a solution of INTERMEDIATE B (2.33 g, 4.81 mmol) in DMF
(60 mL) was added INTERMEDIATE A (0.563 g, 2.075 mmol), cesium
carbonate (2.70 g, 8.30 mmol) and sodium iodide (0.622 g, 4.15
mmol). The mixture was stirred at 40.degree. C. under N.sub.2
balloon for 13 h. LCMS showed the reaction was completed. Then the
mixture was diluted with EtOAc (100 mL) and washed with water
(3.times.100 mL), followed by brine (20 mL). The mixture was
concentrated in vacuo and the crude product was purified by
gradient elution on SiO.sub.2 (ISCO, 120-g RediSep Gold column, 0
to 10% CH.sub.2Cl.sub.2/MeOH over 60 min, dry-loaded) to provide
Int5A_PEAK1 (the firsted eluting isomer) and Int5A_PEAK2 (the
second eluting isomer). Int5A_PEAK1: NMR (400 MHz, CDCl.sub.3)
.delta.=7.55-7.65 (m, 6H), 7.28-7.53 (m, 10H), 6.42 (d, J=7.9 Hz,
1H), 5.52 (d, J=10.5 Hz, 1H), 5.18 (d, J=10.1 Hz, 1H), 4.88 (br d,
J=12.3 Hz, 1H), 4.67 (d, J=14.0 Hz, 1H), 4.25 (br s, 1H), 4.04-4.15
(m, 1H), 2.98-3.15 (m, 2H), 2.06-2.20 (m, 1H), 1.61 (br d, J=8.8
Hz, 4H), 1.13 (s, 9H). LRMS (M+H).sup.+: 594.3. Int5A_PEAK2:
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta.=7.58-7.70 (m, 7H),
7.29-7.42 (m, 9H), 6.37 (d, J=7.5 Hz, 1H), 5.52 (d, J=10.1 Hz, 1H),
5.20-5.29 (m, 2H), 3.49 (s, 3H), 3.15-3.30 (m, 2H), 2.66-2.76 (m,
1H), 2.01-2.18 (m, 2H), 1.79 (br t, J=10.1 Hz, 2H), 1.62 (br s,
3H), 1.07 (s, 9H). LRMS (M+H).sup.+: 594.3.
Step 2
9-(benzyloxy)-4-hydroxy-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[1,2-b][1,2-
,5]triazecine-8,10-dione
##STR00046##
[0188] To a solution of Int5A_PEAK1 (1.66 g, 2.80 mmol) in THF (30
mL) was added tetrabutylammonium fluoride (6.99 mL, 6.99 mmol)(1 M
in THF) under N.sub.2, and then the mixture was stirred at
30.degree. C. for 2 h. The mixture was concentrated in vacou. The
crude product was purified by reverse phase preparative HPLC
(column: Phenomenex Synergi Max-RP 150 mm*50 mm, 10 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 0%, end B 30%; gradient
time 23 min; flowrate: 120 mL/min) to afford Int5B_PEAK1. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta.=7.76 (d, J=7.5 Hz, 1H), 7.50-7.58
(m, 2H), 7.26-7.37 (m, 4H), 5.44 (d, J=10.1 Hz, 1H), 5.23 (d,
J=10.1 Hz, 1H), 4.11-4.26 (m, 1H), 3.72-3.91 (m, 1H), 3.14-3.20 (m,
4H), 1.53-1.61 (m, 4H). LRMS (M+H).sup.+: 356.1. Int5A_PEAK2 was
used to prepare Int5B_PEAK2 in an analogous manner. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta.=7.79 (br d, J=7.1 Hz, 1H), 7.52 (br
d, J 6.2 Hz, 2H), 7.28-7.38 (m, 4H), 5.33-5.48 (m, 1H), 5.24 (br d,
J=9.7 Hz, 1H), 4.40-4.50 (m, 2H), 4.07-4.17 (m, 1H), 3.97 (s, 4H),
3.12 (br s, 4H), 3.04 (br dd, J=8.4, 4.2 Hz, 3H), 2.17-2.31 (m,
1H), 1.85-2.04 (m, 3H), 1.57 (br s, 4H), 1.52-1.73 (m, 19H),
1.30-1.46 (m, 23H), 0.86-1.10 (m, 34H). LRMS (M+H).sup.+: 356.1
Step 3
9-(benzyloxy)-4-hydroxy-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[1,2-b][1,2-
,5]triazecine-8,10-dione
##STR00047##
[0190] To a solution of Int5B_PEAK1 (2.2 g, 6.19 mmol) in
CH.sub.2Cl.sub.2 (80 mL) was added NBS (2.20 g, 12.38 mmol), and
the mixture was stirred at 30.degree. C. for 2 h. The mixture was
quenched with Na.sub.2SO.sub.3 solution (30 mL), extracted with
EtOAc (3.times.10 mL), the organic layer was dried over
Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated in
vacuo. The residue was purified by gradient elution on SiO.sub.2
(ISCO, 4-g RediSep Gold column, 0 to 10% EtOAc/MeOH over 60 min,
dry-loaded) to provide Int5C_PEAK1. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta.=7.87 (s, 1H), 7.62 (d, J=6.8 Hz, 2H), 7.30-7.41
(m, 3H), 5.50 (d, J=9.8 Hz, 1H), 5.20 (d, J=10.3 Hz, 1H), 4.18-4.25
(m, 2H), 3.57-3.79 (m, 1H), 3.43-3.57 (m, 1H), 3.04-3.20 (m, 2H),
2.88-2.99 (m, 1H), 1.74-2.01 (m, 4H). LRMS (M+H).sup.+: 435.8.
Int5B_PEAK2 was used to prepare Int5C_PEAK2 in an analogous manner.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta.=7.83 (s, 1H), 7.58-7.64
(m, 2H), 7.26-7.38 (m, 3H), 5.50 (d, J=9.9 Hz, 1H), 5.19 (d, J=9.9
Hz, 1H), 4.31 (d, J=14.3 Hz, 1H), 3.48 (s, 1H), 2.86 (ddd, J=14.3,
6.8, 4.2 Hz, 2H), 2.20-2.37 (m, 1H), 1.59 (br d, J=6.8 Hz, 2H).
LRMS (M+H).sup.+: 435.8
Step 4
9-(benzyloxy)-11-bromo-4-methoxy-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[1-
,2-b][1,2,5]triazecine-8,10-dione
##STR00048##
[0192] To a solution of Int5C_PEAK1 (70 mg, 0.16 mmol) in DMF (2
mL), cooled to 0.degree. C., was added sodium hydride (12.89 mg,
0.32 mmol) and iodomethane (0.020 mL, 0.32 mmol). The mixture was
stirred under N.sub.2 for 2 h at 30.degree. C. The mixture was
quenched with 1N HCl (0.5 mL), diluted with MeOH (2 mL), and
purified by reverse phase preparative HPLC (Column: Phenomenex
Synergi C18 150 mm*30 mm, 4 .mu.m; Conditions: water (0.1%
TFA)/MeCN; begin B 24%, end B 47%; gradient time 10 min; flowrate:
25 mL/min) to afford Int5D_PEAK1. LRMS (M+H).sup.+: 448.1.
Int5C_PEAK2 was used to prepare Int5D_PEAK2 in an analogous manner.
LRMS (M+H).sup.+: 448.1.
[0193] Step 5:
9-(benzyloxy)-4-methoxy-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-
-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00049##
[0194] To a solution of Int5D_PEAK1 (42 mg, 0.094 mmol) in DMSO (1
mL) was added (2,4,6-trifluorophenyl)methanamine (0.023 mL, 0.19
mmol), N-ethyl-N-isopropylpropan-2-amine (0.082 mL, 0.47 mmol) and
Pd(PPh.sub.3).sub.4 (54.1 mg, 0.047 mmol), and the mixture was
degassed under vacuum and purged with CO (3.times.). The mixture
was stirred at 85.degree. C. under balloon CO (15 psi) for 3 h. The
mixture was diluted with EtOAc (10 mL), filtered, washed with water
(3.times.10 mL), followed by brine (20 mL). The combined organic
layers was dried over Na.sub.2SO.sub.4 and filtered. The filtrate
was concentrated in vacuo, and the residue was purified by
isocratic elution on SiO.sub.2 (CH.sub.2Cl.sub.2:EtOAc=5:1) to
provide crude product, which was further purified by reverse phase
preparative HPLC (Column: YMC-Actus Pro C18 150 mm*30 mm, 4 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 43%, end B 73%; gradient
time 10 min; flowrate: 40 mL/min) to afford Int5E_PEAK1. LRMS
(M+H).sup.+: 557.2. Int5D_PEAK2 was used to prepare Int5E_PEAK2 in
an analogous manner. LRMS (M+H).sup.+: 557.2
Step 6a
9-(benzyloxy)-4-methoxy-8,10-dioxo-N-(2,4,6-trifluorobenzy)-3,4,5,6,8,10-h-
exahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00050##
[0196] Enantiomers were separated by preparative chiral SFC
(Column: CHIRALPAK-AS 250*50 mm, 10 .mu.m; Conditions: isocratic
CO.sub.2/EtOH (0.1% NH.sub.4OH); begin B 40%, end B 40%
(isocratic); flowrate 45 mL/min) to provide Int5E_PEAK1_Ent1 (PEAK
1, >99.9% ee) and Int5E_PEAK1_Ent2 (PEAK 2, 99.02% ee). LRMS
(M+H).sup.+: 557.2
Step 6b
9-(benzyloxy)-4-methoxy-8,10-dioxo-N-(2,4,6-trifluorobenzy)-3,4,5,6,8,10-h-
exahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00051##
[0198] Enantiomers were separated by preparative chiral SFC
(Column: CHIRALPAK-AS 250*50 mm, 10 .mu.m; Conditions: isocratic
CO.sub.2/EtOH (0.1% NH.sub.4OH); begin B 40%, end B 40%
(isocratic); flowrate 45 mL/min) to provide IntSE_PEAK2_Ent1 (PEAK
1, >99.9% ee) and Int5E_PEAK2_Ent2 (PEAK 2, 99.1% ee). LRMS
(M+H).sup.+: 557.2
Step 7
9-hydroxy-4-methoxy-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexa-
hydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00052## ##STR00053##
[0200] To a solution of Int5E_PEAK1_Ent1 (8.8 mg, 0.016 mmol) in
MeCN (5 mL) was added magnesium bromide (29.1 mg, 0.16 mmol). The
mixture was stirred at 30.degree. C. for 2 h. The mixture was
concentrated in vacuo. The residue was diluted with MeOH (3 mL),
filtered, and the crude product was purified by reverse phase
preparative HPLC (Column: Boston Green ODS 150 mm*30 mm, 5 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 40%, end B 70%; gradient
time 10 min; flowrate: 25 mL/min) to afford 5A. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta.=10.33 (br s, 1H), 8.46 (s, 1H), 6.65 (t,
J=8.2 Hz, 2H), 4.79-4.92 (m, 2H), 4.57-4.74 (m, 2H), 4.22 (br d,
J=14.1 Hz, 1H), 3.74-3.86 (m, 1H), 3.67 (br d, J=6.2 Hz, 1H), 3.41
(s, 3H), 3.03-3.21 (m, 2H), 1.93-2.21 (m, 4H). LRMS (M+H).sup.+:
467.0. Int5E_PEAK1_Ent2 was used to prepare 5B in an analogous
manner. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.=10.34 (br s, 1H),
8.47 (s, 1H), 8.45-8.48 (m, 1H), 6.65 (t, J=8.0 Hz, 2H), 4.79-4.92
(m, 2H), 4.58-4.74 (m, 2H), 4.22 (br d, J=13.0 Hz, 1H), 3.75-3.85
(m, 1H), 3.68 (br s, 1H), 3.41 (s, 3H), 3.04-3.20 (m, 2H),
1.93-2.17 (m, 4H). LRMS (M+H).sup.+: 467.0. IntSE_PEAK2_Ent1 was
used to prepare 5C in an analogous manner. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta.=10.23 (br s, 1H), 8.40 (s, 1H), 6.59 (t, J 8.2
Hz, 2H), 4.54-4.74 (m, 3H), 4.38-4.53 (m, 2H), 3.18-3.29 (m, 6H),
2.94 (br d, J=13.7 Hz, 1H), 2.22 (br s, 2H), 1.94 (s, 2H). MS
(M+H).sup.+: 467.0. Int5E_PEAK2_Ent2 was used to prepare 5D in an
analogous manner. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.=10.30
(br s, 1H), 8.46 (s, 1H), 6.65 (t, J=8.0 Hz, 2H), 4.65 (br dd,
J=10.1, 6.0 Hz, 2H), 4.45-4.58 (m, 2H), 3.30 (s, 6H), 3.00 (dt,
J=14.3, 5.1 Hz, 1H), 2.23-2.37 (m, 2H), 1.93-2.05 (m, 2H). MS
(M+H).sup.+: 467.0.
Example 6
##STR00054##
[0201] Step 1
9-(benzyloxy)-4-methyl-8,10-dioxo-3,4,5,6,8,10-hexahydro-2H-1,7-methanopyr-
ido[1,2-b][1,2,5,8]tetrazecin-4-ium 2,2,2-trifluoroacetate
##STR00055##
[0203] To INTERMEDIATE A (600 mg, 1.212 mmol) in DMF (15 mL) cooled
to 0.degree. C. was added sodium hydride (354 mg, 8.85 mmol), and
the reaction mixture was stirred at the same temperature for 30
minutes. 2-chloro-N-(2-chloroethyl)-N-methylethanamine
hydrochloride (1107 mg, 5.75 mmol) was added, and the reaction
mixture was heated to 60.degree. C. for 90 minutes. The mixture was
cooled to 0.degree. C. and sodium hydride (354 mg, 8.85 mmol) was
added again, and the mixture heated to 60.degree. C. after the
addition for 20 minutes. The mixture was cooled to room temperature
and quenched with 1:1 MeCN/water (3 mL) and then diluted with water
(250 mL). The mixture in water was purified directly by preparative
HPLC (Column: Phenomenex Luna C18 250*50 mm, 10 .mu.m; Conditions:
water (0.1% TFA)/MeCN; begin B 0%, end B 50%; gradient time 25 min;
flowrate 90 mL/min) to provide the title compound. .sup.1H NMR (500
MHz, CD.sub.3OD) .delta.=7.98 (d, J=7 Hz, 1H), 7.52 (m, 2H), 7.37
(m, 3H), 6.64 (d, J=7 Hz, 1H), 5.34 (d, J=10.5 Hz, 1H), 5.21 (d,
J=11 Hz, 1H), 4.98 (d, J=15.5 Hz, 1H), 4.88 (m, 1H), 4.02 (m, 1H),
3.75-3.69 (m, 2H), 3.47-3.33 (m, 3H), 3.01 (s, 3H). LRMS
(M+H).sup.+: 355.4.
Step 2
9-(benzyloxy)-11-iodo-4-methyl-8,10-dioxo-3,4,5,6,8,10-hexahydro-2H-1,7-me-
thanopyrido[1,2-b][1,2,5,8]tetrazecine 4-oxide
2,2,2-trifluoroacetate
##STR00056##
[0205] To a stirred solution of
9-(benzyloxy)-4-methyl-8,10-dioxo-3,4,5,6,8,10-hexahydro-2H-1,7-methanopy-
rido[1,2-b][1,2,5,8]tetrazecin-4-ium 2,2,2-trifluoroacetate (360
mg, 0.769 mmol) in MeOH (7.7 mL) was added mCPBA (265 mg, 1.54
mmol). The reaction mixture stirred for 5 minutes at room
temperature. NIS (432 mg, 1.92 mmol) was then added in one portion,
and the mixture was stirred at 70.degree. C. for 2 h. The mixture
was concentrated. The residue was dissolved in 1:1 MeCN/water (8
mL) and purified by preparative HPLC (Column: Waters XBridge C18
250*30 mm, 5 .mu.m; Conditions: water (0.1% TFA)/MeCN; begin B 5%,
end B 75%; gradient time 20 min; 100% B hold time 1.1 min, flowrate
50 mL/min) to afford the title compound. LRMS (M+H).sup.+:
497.4.
Step 3
9-(benzyloxy)-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-h-
exahydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxamide
##STR00057##
[0207] A mixture of
9-(benzyloxy)-11-iodo-4-methyl-8,10-dioxo-3,4,5,6,8,10-hexahydro-2H-1,7-m-
ethanopyrido[1,2-b][1,2,5,8]tetrazecine 4-oxide
2,2,2-trifluoroacetate (70 mg, 0.12 mmol), Pd(dppf)C1.sub.2 (8.4
mg, 0.011 mmol) and DIEA (0.074 mL, 0.57 mmol) in DMSO (1.1 mL) was
stirred under CO balloon (15 psi) at 80.degree. C. for 4 h. The
mixture was diluted with DMSO (2 mL) and purified directly by
preparative HPLC (Column: Waters Sunfire C18 150*30 mm, 5 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 15%, end B 85%; gradient
time 20 min; 100% B hold time 1.1 min, flowrate 40 mL/min) and
fractions containing product were neutralized with NaHCO.sub.3
(aq), extracted with DCM (2.times.30 mL) and concentrated to afford
the title compound. LRMS (M+H).sup.+: 542.5.
Step 4
9-(benzyloxy)-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-h-
exahydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxamide
##STR00058##
[0209] Enantiomers of
9-(benzyloxy)-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10--
hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxamide
(29 mg, 0.054 mmol) were separated by preparative chiral SFC
(Column: AS-H column 250*20 mm, 5 .mu.m; Conditions: isocratic
CO.sub.2/MeOH (0.1% DEA); begin B 40%, end B 40% (isocratic);
flowrate 60 mL/min; peak 1 rt=1.55 min, peak 2 rt=2.56 min) to
provide
(1R)-9-(benzyloxy)-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,-
8,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxam-
ide (PEAK 1, >99.5% ee) and
(1S)-9-(benzyloxy)-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,-
8,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxam-
ide (PEAK 2, >99.5% ee). LRMS (M+H).sup.+: 542.5.
Step 5
9-hydroxy-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexah-
ydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxamide
2,2,2-trifluoroacetate
##STR00059##
[0211] To a solution of PEAK 1 of
9-(benzyloxy)-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10--
hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxamide
(10 mg, 0.018 mmol) in DCM (0.5 mL) was added TFA (0.5 mL, 6.49
mmol) and the mixture was stirred at 23.degree. C. for 2 h. LCMS
showed the reaction was completed. The mixture was concentrated.
The residue was dissolved in 1:1 MeCN/water (2 mL) and purified by
prep HPLC HPLC (Column: Waters Sunfire C18 150*30 mm, 5 .mu.m;
Conditions: water (0.1% TFA)/MeCN; begin B 15%, end B 85%; gradient
time 20 min; 100% B hold time 1.1 min, flowrate 40 mL/min) to give
9-hydroxy-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10-hexa-
hydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxamide
2,2,2-trifluoroacetate 6A. .sup.1H NMR (600 MHz, CD.sub.3OD)
.delta.=10.43 (s, 1H), 8.47 (s, 1H), 6.89 (m, 2H), 5.14 (d, J=14.4
Hz, 1H), 5.03 (d, J=14.4 Hz, 1H), 4.65 (m, 2H), 4.58 (d, J=16.2 Hz,
1H), 3.99 (m, 1H), 3.76 (m, 1H), 3.63 (brs, 2H), 3.50 (m, 1H), 3.37
(m, 2H), 2.95 (s, 3H). LRMS (M+H).sup.+: 452.5. PEAK 2 of
9-(benzyloxy)-4-methyl-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10--
hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5,8]tetrazecine-11-carboxamide
(10 mg, 0.018 mmol) from the preceding chiral separation was
subjected to the same reaction conditions to afford 6B. .sup.1H NMR
(600 MHz, CD.sub.3OD) .delta.=10.43 (s, 1H), 8.47 (s, 1H), 6.89 (m,
2H), 5.14 (d, J=14.4 Hz, 1H), 5.03 (d, J=14.4 Hz, 1H), 4.65 (m,
2H), 4.58 (d, J=16.2 Hz, 1H), 3.99 (m, 1H), 3.76 (m, 1H), 3.63
(brs, 2H), 3.50 (m, 1H), 3.37 (m, 2H), 2.95 (s, 3H). LRMS
(M+H).sup.+: 452.5.
Example 7
##STR00060##
[0212] Step 1
9-(benzyloxy)-11-bromo-5,6-dihydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triaz-
ecine-4,8,10(3H)-trione
##STR00061##
[0214] To a solution of
9-(benzyloxy)-11-bromo-4-hydroxy-3,4,5,6-tetrahydro-2H-1,7-methanopyrido[-
1,2-b][1,2,5]triazecine-8,10-dione (60 mg, 0.14 mmol) in
CH.sub.2Cl.sub.2 (5 mL) was added DMP (58.6 mg, 0.14 mmol) at
0.degree. C. The mixture was stirred at 0.degree. C. for 2 h. LCMS
showed starting material was consumed and the desired MS was
observed, then the reaction was quenched with MeOH, diluted with
water (5 mL) and extracted with DCM/MeOH (10/1) (20 mL.times.3).
The combined organic phase was washed with brine (10 mL), dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo, the
crude product was purified by normal phase chromatography (ISCO, 4
g RediSep Gold column, 0-10% EtOAc/MeOH, 30 min, dry loaded) to
afford
9-(benzyloxy)-11-bromo-5,6-dihydro-2H-1,7-methanopyrido[1,2-b][1,2,5]tria-
zecine-4,8,10(3H)-trione. .sup.1H NMR (CHLOROFORM-d) .delta.: 7.80
(s, 1H), 7.54 (br d, J=6.6 Hz, 2H), 7.20-7.32 (m, 3H), 5.52 (d,
J=10.1 Hz, 1H), 5.17 (d, J=10.1 Hz, 1H), 4.53-4.72 (m, 2H), 3.99
(d, J=15.2 Hz, 1H), 3.32-3.70 (m, 3H), 3.00 (br s, 1H), 2.72 (br s,
1H), 2.45 (br s, 1H), 2.28 (br d, J=11.0 Hz, 1H). MS (ESI) m/z:
432.1, 434.1 [M+H.sup.+].
Step 2
9-(benzyloxy)-11-bromo-4,4-difluoro-3,4,5,6-tetrahydro-2H-1,7-methanopyrid-
o[1,2-b][1,2,5]triazecine-8,10-dione
##STR00062##
[0216] To a solution of
9-(benzyloxy)-11-bromo-5,6-dihydro-2H-1,7-methanopyrido[1,2-b][1,2,5]tria-
zecine-4,8,10(3H)-trione (45 mg, 0.104 mmol) in
C1CH.sub.2CH.sub.2C1 (5 mL) was added BAST (0.096 ml, 0.521 mmol)
at 0.degree. C., the mixture was stirred at 28.degree. C. for 4 h,
LCMS showed there was .about.20% conversion, then the reaction
mixture was stirred at 28.degree. C. for another 24 h. LCMS showed
there was .about.80% conversion. The reaction was quenched with aq
NaHCO.sub.3 (0.5 mL) at 0.degree. C., diluted with water (5 mL) and
extracted with DCM/MeOH=10/1 (10 mL.times.3), the combined organic
was dried over Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by preparative TLC plate eluting
with 6.7% MeOH/DCM to give
9-(benzyloxy)-11-bromo-4,4-difluoro-3,4,5,6-tetrahydro-2H-1,7-methanopyri-
do[1,2-b][1,2,5]triazecine-8,10-dione. .sup.1H NMR (CHLOROFORM-d)
.delta.: 7.73-7.88 (m, 1H), 7.48-7.59 (m, 2H), 7.22-7.31 (m, 3H),
5.49 (d, J=10.1 Hz, 1H), 5.07-5.18 (m, 1H), 4.70-4.87 (m, 1H),
4.24-4.39 (m, 2H), 3.25-3.74 (m, 3H), 3.01 (ddd, J=14.4, 10.7, 3.7
Hz, 1H), 1.98-2.41 (m, 3H). MS (ESI) m/z: 454.1, 456.1
[M+H.sup.+].
Step 3
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,-
10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00063##
[0218] To a solution of
9-(benzyloxy)-11-bromo-4,4-difluoro-3,4,5,6-tetrahydro-2H-1,7-methanopyri-
do[1,2-b][1,2,5]triazecine-8,10-dione (20 mg, 0.044 mmol) in DMSO
(2 mL) was added (2,4,6-trifluorophenyl)methanamine (21.28 mg,
0.132 mmol), Pd(Ph.sub.3P).sub.4 (5.09 mg, 4.40 .mu.mol) and DIEA
(0.038 mL, 0.22 mmol), then the reaction mixture was degassed and
purged with CO for three times. The mixture was stirred at
85.degree. C. for 2 h under CO balloon. LCMS showed the reaction
was completed. The reaction mixture was diluted with EtOAc (10 mL),
filtered and washed with HCl (5 mL, 1M/L). The HCl layers was
extracted with EtOAc (10 mL.times.3), the combined the organic
layer was dried over Na.sub.2SO.sub.4 and filtered, concentrated in
vacuo, the residue was purified by preparative TLC plate eluting
with 100% EtOAc to afford
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8-
,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide.
.sup.1H NMR (CHLOROFORM-d) .delta.: 10.24 (br s, 1H), 8.43-8.57 (m,
1H), 7.43-7.57 (m, 2H), 7.23-7.35 (m, 3H), 6.59-6.64 (m, 1H), 6.59
(q, J=8.2 Hz, 2H), 5.44 (d, J=10.4 Hz, 1H), 5.16 (d, J=9.9 Hz, 1H),
4.52-4.80 (m, 3H), 4.24-4.38 (m, 3H), 3.65 (s, 1H), 3.33-3.52 (m,
1H), 3.04 (br d, J=11.5 Hz, 1H), 2.02-2.75 (m, 2H). MS (ESI) m/z:
563.0 [M+H.sup.+].
Step 4
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,-
10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00064##
[0220] Resolution to the enantiomer
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8-
,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
(20 mg, 0.036 mmol) was accomplished with SFC (Column: DAICEL
CHIRALPAK AS (250 mm*50 mm, 10 .mu.m); Mobile phase: 35% Base-EtOH
(contained 0.1% NH.sub.3H.sub.2O) in CO.sub.2; Flow rate: 50
mL/min; injection: 150) to give
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,-
5,6,8,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxa-
mide as the first eluting isomer (PEAK 1) and
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8-
,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
as the second eluting isomer (PEAK 2). MS (ESI) m/z: 563.0
[M+H.sup.+].
Step 5
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,-
10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
##STR00065##
[0222] To a solution of PEAK 1 of
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8-
,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
(9 mg, 0.016 mmol) in MeCN (3 mL) was added magnesium bromide
(14.73 mg, 0.080 mmol). The mixture was stirred at 22.degree. C.
for 2 h. LCMS showed the reaction was completed. The mixture was
concentrated in vacuo and the residue was diluted with MeOH (2 mL),
filtrated, the crude product was purified by preparative HPLC
(column: Boston Green ODS 150 mm*30 mm, 5 .mu.m; condition: water
(0.1% TFA)-MeCN begin B 37, end B 67; gradient time (min): 10; 100%
B Hold time (min): 2; FlowRate (mL/min)L 25) to give
4,4-difluoro-9-hydroxy-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8,10--
hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
7A and which was Co-evaporated with toluene to afford the free
base. .sup.1H NMR (CHLOROFORM-d) .delta.: 10.11 (br s, 1H), 8.42
(s, 1H), 6.59 (t, J=8.2 Hz, 2H), 4.90 (br d, J=14.1 Hz, 1H),
4.52-4.68 (m, 2H), 4.25-4.46 (m, 2H), 3.72 (br d, J=11.7 Hz, 1H),
3.28-3.47 (m, 1H), 3.03-3.20 (m, 1H), 2.50-2.75 (m, 1H), 2.02-2.38
(m, 3H). MS (ESI) m/z: 472.9 [M+H.sup.+]PEAK 2 of
9-(benzyloxy)-4,4-difluoro-8,10-dioxo-N-(2,4,6-trifluorobenzyl)-3,4,5,6,8-
,10-hexahydro-2H-1,7-methanopyrido[1,2-b][1,2,5]triazecine-11-carboxamide
(9 mg, 0.016 mmol) from the preceding chiral separation was
subjected to the same reaction conditions to afford 7B. .sup.1H NMR
(CHLOROFORM-d) .delta.: 10.12 (br s, 1H), 8.42 (s, 1H), 6.59 (t,
J=8.0 Hz, 2H), 4.90 (br d, J=13.5 Hz, 1H), 4.52-4.69 (m, 2H),
4.34-4.48 (m, 2H), 3.74 (br s, 1H), 3.39 (br s, 1H), 3.14 (br t,
J=10.7 Hz, 1H), 2.49-2.77 (m, 1H), 2.06-2.31 (m, 3H). MS (ESI) m/z:
472.9 [M+H.sup.+]
[0223] The compounds in Table 1 were prepared in an analogous
fashion to that described in the examples above. The isomers were
separated by either preparative HPLC, preparative chiral HPLC, or
preparative chiral SFC. Assigned stereochemistry is done so by
analogy to compounds 1A and 1B. Unknown and unassigned
strerochemistry is denoted by an asterisk (*).
TABLE-US-00002 TABLE 1 Preparative LRMS Ex. No. Structure Method(s)
Used (ESI) [M + 1].sup.+ 8A ##STR00066## Example 1 419 8B
##STR00067## Example 1 419 9A ##STR00068## Example 1 435 9B
##STR00069## Example 1 435 10A ##STR00070## Example 1 453 10B
##STR00071## Example 1 453 11A ##STR00072## Example 1 453 11B
##STR00073## Example 1 453 12A ##STR00074## Example 3 467 12B
##STR00075## Example 3 467 13A ##STR00076## Example 4 437 13B
##STR00077## Example 4 437 14A ##STR00078## Example 4 439 14B
##STR00079## Example 4 439 15A ##STR00080## Example 4 455 15B
##STR00081## Example 4 455 16A ##STR00082## Example 4 455 16B
##STR00083## Example 4 455 17A ##STR00084## Example 5 453 17B
##STR00085## Example 5 453 17C ##STR00086## Example 5 453 17D
##STR00087## Example 5 453 18A ##STR00088## Example 5 465 18B
##STR00089## Example 5 465 18C ##STR00090## Example 5 465 18D
##STR00091## Example 5 465 19A ##STR00092## Example 5 483 19B
##STR00093## Example 5 483 19C ##STR00094## Example 5 483 19D
##STR00095## Example 5 483 20A ##STR00096## Example 5 449 20B
##STR00097## Example 5 449 20C ##STR00098## Example 5 449 20D
##STR00099## Example 5 449 21A ##STR00100## Example 5 483 21B
##STR00101## Example 5 483 21C ##STR00102## Example 5 483 21D
##STR00103## Example 5 483 22A ##STR00104## Example 5 449 22B
##STR00105## Example 5 449 22C ##STR00106## Example 5 449 22D
##STR00107## Example 5 449 23A ##STR00108## Example 5 449 23B
##STR00109## Example 5 449 23C ##STR00110## Example 5 449 23D
##STR00111## Example 5 449 24A ##STR00112## Example 6 450 24B
##STR00113## Example 6 450 25A ##STR00114## Example 6 468 25B
##STR00115## Example 6 468 26A ##STR00116## Example 7 455 26B
##STR00117## Example 7 455
Assessing Antiviral Potency in a Multiple Round HIV-1 Infection
Assay
[0224] The antiviral activity of the Examples herein was assessed
in an assay that measures the rate of replication of HIV in cell
culture, and performed according to the following procedure. HIV-1
replication was monitored using MT4-gag-GFP clone D3 (hereafter
designated MT4-GFP), which are MT-4 cells modified to harbor a GFP
reporter gene, the expression of which is dependent on the HIV-1
expressed proteins t at and rev. Productive infection of an MT4-GFP
cell with HIV-1 results in GFP expression approximately 24 h
post-infection. MT4-GFP cells were maintained at 37.degree. C./5%
CO.sub.2/90% relative humidity in RPMI 1640 supplemented with 10%
fetal bovine serum, 100 U/ml penicillin/streptomycin, and 400
.mu.g/ml G418 to maintain the reporter gene. For infections,
MT4-GFP cells were placed in the same medium lacking G418 and
infected overnight with HIV-1 (H9/IIIB strain) virus at an
approximate multiplicity of infection of 0.01 in the same
incubation conditions. Cells were then washed and re-suspended in
either RPMI 1640 at 2.times.10.sup.5 cells/mL (0% NHS condition) or
100% normal human serum (NHS) at 2.times.10.sup.5 cells/mL (100%
NHS condition). Compound plates were prepared by dispensing
compounds dissolved in DMSO into wells of 384 well
poly-D-lysine-coated plates (0.2 .mu.l/well) using an ECHO acoustic
dispenser. Each compound was tested in a 10-point serial 3-fold
dilution (typical final concentrations: 1050 nM-0.05 nM for 0% NHS
condition or 42 .mu.M-2.13 nM for 100% NHS condition). Controls
included no inhibitor (DMSO only) and a combination of three
antiviral agents (efavirenz, indinavir, an in-house integrase
strand transfer inhibitor at final concentrations of 4 .mu.M each).
Cells were added (504/well) to compound plates and the infected
cells were maintained at 37.degree. C./5% CO.sub.2/90% relative
humidity.
[0225] Infected cells were quantified at two time points, .about.48
h and .about.72 h post-infection, by counting the number of green
cells in each well using an Acumen eX3 scanner. The increase in the
number of green cells over .about.24 h period gives the
reproductive ratio, RO, which is typically 5-15 and has been shown
experimentally to be in logarithmic phase (data not shown).
Inhibition of RO is calculated for each well, and IC.sub.50s
determined by non-linear 4-parameter curve fitting. Assay IC.sub.50
results are shown in the table below.
TABLE-US-00003 Viking, IC.sub.50 Viking, IC.sub.50 Example (0% NHS)
(nM) (100% NHS) (nM) 1A 1.98 2559 1B 1.00 94.7 2A 4.04 4164 2B 1.53
234.4 3A 2.66 589 3B 5.23 1885 4A 3.45 9518 4B 1.78 196.7 5A 3.20
67.2 5B 1.93 124 5C 1.68 62.8 5D 5.32 543.5 6A 4.64 169.2 6B 2.40
80.2 7A 6.1 4462 7B 2.16 73.9 8A 1.03 1567 8B 0.86 122.2 9A 2.57
5621 9B 1.15 196 10A 1.89 8581 10B 1.65 225 11A 3.45 12510 11B 2.09
357.8 12A 14.28 10710 12B 3.21 427.9 13A 2.74 3364 13B 2.19 145.7
14A 5.40 4213 14B 1.66 156.6 15A 3.70 10660 15B 1.61 224.4 16A 2.7
12280 16B 1.2 247 17A 66.1 1266 17B 7.6 275 17C 184.4 5919 17D 25.6
717 18A 3.1 279 18B 2.5 435 18C 3.2 1873 18D 1.5 230 19A 1.8 116
19B 5.3 1875 19C 3.5 245 19D 1.5 300 20A 3.3 88 20B 2.8 371 20C 7.5
606 20D 2.1 212 21A 2.5 2522 21B 0.9 207 21C 1.7 311 21D 2.0 472
22A 8.2 333 22B 1.6 161 22C 2.0 88 22D 1.4 311 23A 5.8 728 23B 2.1
314 23C 4.3 83 23D 1.9 567 24A 5.1 665 24B 3.5 485 25A 4.5 1001 25B
2.1 139 26A 6.6 869 26B 3.2 91
Treatment or Prevention of HIV Infection
[0226] The Tricyclic Heterocycle Compounds may be useful in the
inhibition of HIV, the inhibition of HIV integrase, the treatment
of HIV infection and/or reduction of the likelihood or severity of
symptoms of HIV infection and the inhibition of HIV viral
replication and/or HIV viral production in a cell-based system. For
example, the Tricyclic Heterocycle Compounds may be useful in
treating infection by HIV after suspected past exposure to HIV by
such means as blood transfusion, exchange of body fluids, bites,
accidental needle stick, or exposure to subject blood during
surgery or other medical procedures.
[0227] Accordingly, in one embodiment, the invention provides
methods for treating HIV infection in a subject, the methods
comprising administering to the subject an effective amount of at
least one Tricyclic Heterocycle Compound or a pharmaceutically
acceptable salt or prodrug thereof. In a specific embodiment, the
amount administered is effective to treat or prevent infection by
HIV in the subject. In another specific embodiment, the amount
administered is effective to inhibit HIV viral replication and/or
viral production in the subject. In one embodiment, the HIV
infection has progressed to AIDS.
[0228] The Tricyclic Heterocycle Compounds are also useful in the
preparation and execution of screening assays for antiviral
compounds. For example the Tricyclic Heterocycle Compounds may be
useful for identifying resistant HIV cell lines harboring
mutations, which are excellent screening tools for more powerful
antiviral compounds. Furthermore, the Tricyclic Heterocycle
Compounds may be useful in establishing or determining the binding
site of other antivirals to the HIV Integrase.
[0229] The compositions and combinations of the present invention
may be useful for treating a subject suffering from infection
related to any HIV genotype.
Combination Therapy
[0230] In another embodiment, the present methods for treating or
preventing HIV infection can further comprise the administration of
one or more additional therapeutic agents which are not Tricyclic
Heterocycle Compounds.
[0231] In one embodiment, the additional therapeutic agent is an
antiviral agent.
[0232] In another embodiment, the additional therapeutic agent is
an immunomodulatory agent, such as an immunosuppressive agent.
[0233] Accordingly, in one embodiment, the present invention
provides methods for treating a viral infection in a subject, the
method comprising administering to the subject: (i) at least one
Tricyclic Heterocycle Compound (which may include two or more
different Tricyclic Heterocycle Compounds), or a pharmaceutically
acceptable salt or prodrug thereof, and (ii) at least one
additional therapeutic agent that is other than a Tricyclic
Heterocycle Compound, wherein the amounts administered are together
effective to treat or prevent a viral infection.
[0234] When administering a combination therapy of the invention to
a subject, therapeutic agents in the combination, or a
pharmaceutical composition or compositions comprising therapeutic
agents, may be administered in any order such as, for example,
sequentially, concurrently, together, simultaneously and the like.
The amounts of the various actives in such combination therapy may
be different amounts (different dosage amounts) or same amounts
(same dosage amounts). Thus, for non-limiting illustration
purposes, a Tricyclic Heterocycle Compound and an additional
therapeutic agent may be present in fixed amounts (dosage amounts)
in a single dosage unit (e.g., a capsule, a tablet and the
like).
[0235] In one embodiment, at least one Tricyclic Heterocycle
Compound is administered during a time when the additional
therapeutic agent(s) exert their prophylactic or therapeutic
effect, or vice versa.
[0236] In another embodiment, at least one Tricyclic Heterocycle
Compound and the additional therapeutic agent(s) are administered
in doses commonly employed when such agents are used as monotherapy
for treating a viral infection.
[0237] In another embodiment, at least one Tricyclic Heterocycle
Compound and the additional therapeutic agent(s) are administered
in doses lower than the doses commonly employed when such agents
are used as monotherapy for treating a viral infection.
[0238] In still another embodiment, at least one Tricyclic
Heterocycle Compound and the additional therapeutic agent(s) act
synergistically and are administered in doses lower than the doses
commonly employed when such agents are used as monotherapy for
treating a viral infection.
[0239] In one embodiment, at least one Tricyclic Heterocycle
Compound and the additional therapeutic agent(s) are present in the
same composition. In one embodiment, this composition is suitable
for oral administration. In another embodiment, this composition is
suitable for intravenous administration. In another embodiment,
this composition is suitable for subcutaneous administration. In
still another embodiment, this composition is suitable for
parenteral administration.
[0240] Viral infections and virus-related disorders that may be
treated or prevented using the combination therapy methods of the
present invention include, but are not limited to, those listed
above.
[0241] In one embodiment, the viral infection is HIV infection.
[0242] In another embodiment, the viral infection is AIDS.
[0243] The at least one Tricyclic Heterocycle Compound and the
additional therapeutic agent(s) can act additively or
synergistically. A synergistic combination may allow the use of
lower dosages of one or more agents and/or less frequent
administration of one or more agents of a combination therapy. A
lower dosage or less frequent administration of one or more agents
may lower toxicity of therapy without reducing the efficacy of
therapy.
[0244] In one embodiment, the administration of at least one
Tricyclic Heterocycle Compound and the additional therapeutic
agent(s) may inhibit the resistance of a viral infection to these
agents.
[0245] As noted above, the present invention is also directed to
use of a compound of Formula I with one or more anti-HIV agents. An
"anti-HIV agent" is any agent which is directly or indirectly
effective in the inhibition of HIV reverse transcriptase or another
enzyme required for HIV replication or infection, the treatment or
prophylaxis of HIV infection, and/or the treatment, prophylaxis or
delay in the onset or progression of AIDS. It is understood that an
anti-HIV agent is effective in treating, preventing, or delaying
the onset or progression of HIV infection or AIDS and/or diseases
or conditions arising therefrom or associated therewith. For
example, the compounds of this invention may be effectively
administered, whether at periods of pre-exposure and/or
post-exposure, in combination with effective amounts of one or more
anti-HIV agents selected from HIV antiviral agents,
immunomodulators, antiinfectives, or vaccines useful for treating
HIV infection or AIDS. Suitable HIV antivirals for use in
combination with the compounds of the present invention include,
for example, those listed in Table A as follows:
TABLE-US-00004 TABLE A Name Type abacavir, ABC, Ziagen .RTM. nRTI
abacavir + lamivudine, Epzicom .RTM. nRTI abacavir + lamivudine +
zidovudine, Trizivir .RTM. nRTI amprenavir, Agenerase .RTM. PI
atazanavir, Reyataz .RTM. PI AZT, zidovudine, azidothymidine,
Retrovir .RTM. nRTI darunavir, Prezista .RTM. PI ddC, zalcitabine,
dideoxycytidine, Hivid .RTM. nRTI ddI, didanosine, dideoxyinosine,
Videx .RTM. nRTI ddI (enteric coated), Videx EC .RTM. nRTI
delavirdine, DLV, Rescriptor .RTM. nnRTI dolutegravir, Tivicay
.RTM. II doravirine, Delstrigo .TM. nnRTI doravirine + lamivudine +
tenofovir disoproxil nnRTI + nRTI + fumarate, Pifeltro .TM. nRTI
efavirenz, EFV, Sustiva .RTM., Stocrin .RTM. nnRTI efavirenz +
emtricitabine + tenofovir DF, Atripla .RTM. nnRTI + nRTI EFdA
(4'-ethynyl-2-fluoro-2'-deoxyadenosine) nRTI emtricitabine, FTC,
Emtriva .RTM. nRTI emtricitabine + tenofovir DF, Truvada .RTM. nRTI
emvirine, Coactinon .RTM. nnRTI enfuvirtide, Fuzeon .RTM. FI
enteric coated didanosine, Videx EC .RTM. nRTI etravirine, TMC-125
nnRTI fosamprenavir calcium, Lexiva .RTM. PI indinavir, Crixivan
.RTM. PI lamivudine, 3TC, Epivir .RTM. nRTI lamivudine +
zidovudine, Combivir .RTM. nRTI lopinavir PI lopinavir + ritonavir,
Kaletra .RTM. PI maraviroc, Selzentry .RTM. EI nelfinavir, Viracept
.RTM. PI nevirapine, NVP, Viramune .RTM. nnRTI rilpivirine, TMC-278
nnRTI ritonavir, Norvir .RTM. PI saquinavir, Invirase .RTM.,
Fortovase .RTM. PI stavudine, d4T, didehydrodeoxythymidine, Zerit
.RTM. nRTI tenofovir DF (DF = disoproxil fumarate), TDF, nRTI
Viread .RTM. tipranavir, Aptivus .RTM. PI EI = entry inhibitor; FI
= fusion inhibitor; PI = protease inhibitor; nRTI = nucleoside
reverse transcriptase inhibitor; II = integrase inhibitor; nnRTI =
non-nucleoside reverse transcriptase inhibitor. Some of the drugs
listed in the table are used in a salt form; e.g., abacavir
sulfate, indinavir sulfate, atazanavir sulfate, nelfinavir
mesylate.
[0246] In one embodiment, one or more anti-HIV drugs are selected
from, lamivudine, abacavir, ritonavir, darunavir, atazanavir,
emtricitabine, tenofovir, rilpivirine and lopinavir.
[0247] In another embodiment, the compound of formula (I) is used
in combination with lamivudine.
[0248] In still another embodiment, the compound of formula (I) is
used in combination atazanavir.
[0249] In another embodiment, the compound of formula (I) is used
in combination with darunavir.
[0250] In another embodiment, the compound of formula (I) is used
in combination with rilpivirine.
[0251] In one embodiment, the compound of formula (I) is used in
combination with lamivudine and abacavir.
[0252] In another embodiment, the compound of formula (I) is used
in combination with darunavir.
[0253] In another embodiment, the compound of formula (I) is used
in combination with emtricitabine and tenofovir.
[0254] In still another embodiment, the compound of formula (I) is
used in combination atazanavir.
[0255] In another embodiment, the compound of formula (I) is used
in combination with ritonavir and lopinavir.
[0256] In one embodiment, the compound of formula (I) is used in
combination with abacavir and lamivudine.
[0257] In another embodiment, the compound of formula (I) is used
in combination with lopinavir and ritonavir.
[0258] In one embodiment, the present invention provides
pharmaceutical compositions comprising (i) a compound of formula
(I) or a pharmaceutically acceptable salt or prodrug thereof; (ii)
a pharmaceutically acceptable carrier; and (iii) one or more
additional anti-HIV agents selected from lamivudine, abacavir,
ritonavir and lopinavir, or a pharmaceutically acceptable salt or
prodrug thereof, wherein the amounts present of components (i) and
(iii) are together effective for the treatment or prophylaxis of
infection by HIV or for the treatment, prophylaxis, or delay in the
onset or progression of AIDS in the subject in need thereof.
[0259] In another embodiment, the present invention provides a
method for the treatment or prophylaxis of infection by HIV or for
the treatment, prophylaxis, or delay in the onset or progression of
AIDS in a subject in need thereof, which comprises administering to
the subject (i) a compound of formula (I) or a pharmaceutically
acceptable salt or prodrug thereof and (ii) one or more additional
anti-HIV agents selected from lamivudine, abacavir, ritonavir and
lopinavir, or a pharmaceutically acceptable salt or prodrug
thereof, wherein the amounts administered of components (i) and
(ii) are together effective for the treatment or prophylaxis of
infection by HIV or for the treatment, prophylaxis, or delay in the
onset or progression of AIDS in the subject in need thereof.
[0260] It is understood that the scope of combinations of the
compounds of this invention with anti-HIV agents is not limited to
the HIV antivirals listed in Table A, but includes in principle any
combination with any pharmaceutical composition useful for the
treatment or prophylaxis of AIDS. The HIV antiviral agents and
other agents will typically be employed in these combinations in
their conventional dosage ranges and regimens as reported in the
art, including, for example, the dosages described in the
Physicians' Desk Reference, Thomson PDR, Thomson PDR, 57.sup.th
edition (2003), the 58.sup.th edition (2004), the 59.sup.th edition
(2005), and the like. The dosage ranges for a compound of the
invention in these combinations are the same as those set forth
above.
[0261] The doses and dosage regimen of the other agents used in the
combination therapies of the present invention for the treatment or
prevention of HIV infection may be determined by the attending
clinician, taking into consideration the approved doses and dosage
regimen in the package insert; the age, sex and general health of
the subject; and the type and severity of the viral infection or
related disease or disorder. When administered in combination, the
Tricyclic Heterocycle Compound(s) and the other agent(s) may be
administered simultaneously (i.e., in the same composition or in
separate compositions one right after the other) or sequentially.
This is particularly useful when the components of the combination
are given on different dosing schedules, e.g., one component is
administered once daily and another component is administered every
six hours, or when the pharmaceutical compositions are different,
e.g., one is a tablet and one is a capsule. A kit comprising the
separate dosage forms is therefore advantageous.
Compositions and Administration
[0262] When administered to a subject, the Tricyclic Heterocycle
Compounds may be administered as a component of a composition that
comprises a pharmaceutically acceptable carrier or vehicle. The
present invention provides pharmaceutical compositions comprising
an effective amount of at least one Tricyclic Heterocycle Compound
and a pharmaceutically acceptable carrier. In the pharmaceutical
compositions and methods of the present invention, the active
ingredients will typically be administered in admixture with
suitable carrier materials suitably selected with respect to the
intended form of administration, i.e., oral tablets, capsules
(either solid-filled, semi-solid filled or liquid filled), powders
for constitution, oral gels, elixirs, dispersible granules, syrups,
suspensions, and the like, and consistent with conventional
pharmaceutical practices. For example, for oral administration in
the form of tablets or capsules, the active drug component may be
combined with any oral non-toxic pharmaceutically acceptable inert
carrier, such as lactose, starch, sucrose, cellulose, magnesium
stearate, dicalcium phosphate, calcium sulfate, talc, mannitol,
ethyl alcohol (liquid forms) and the like. Solid form preparations
include powders, tablets, dispersible granules, capsules, cachets
and suppositories. Powders and tablets may be comprised of from
about 0.5 to about 95 percent inventive composition. Tablets,
powders, cachets and capsules may be used as solid dosage forms
suitable for oral administration.
[0263] Moreover, when desired or needed, suitable binders,
lubricants, disintegrating agents and coloring agents may also be
incorporated in the mixture. Suitable binders include starch,
gelatin, natural sugars, corn sweeteners, natural and synthetic
gums such as acacia, sodium alginate, carboxymethylcellulose,
polyethylene glycol and waxes. Among the lubricants there may be
mentioned for use in these dosage forms, boric acid, sodium
benzoate, sodium acetate, sodium chloride, and the like.
Disintegrants include starch, methylcellulose, guar gum, and the
like. Sweetening and flavoring agents and preservatives may also be
included where appropriate.
[0264] Liquid form preparations include solutions, suspensions and
emulsions and may include water or water-propylene glycol solutions
for parenteral injection.
[0265] Liquid form preparations may also include solutions for
intranasal administration.
[0266] Also included are solid form preparations which are intended
to be converted, shortly before use, to liquid form preparations
for either oral or parenteral administration. Such liquid forms
include solutions, suspensions and emulsions.
[0267] For preparing suppositories, a low melting wax such as a
mixture of fatty acid glycerides or cocoa butter is first melted,
and the active ingredient is dispersed homogeneously therein as by
stirring. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool and thereby solidify.
[0268] Additionally, the compositions of the present invention may
be formulated in sustained release form to provide the rate
controlled release of any one or more of the components or active
ingredients to optimize therapeutic effects, i.e., antiviral
activity and the like. Suitable dosage forms for sustained release
include layered tablets containing layers of varying disintegration
rates or controlled release polymeric matrices impregnated with the
active components and shaped in tablet form or capsules containing
such impregnated or encapsulated porous polymeric matrices.
[0269] In one embodiment, the one or more Tricyclic Heterocycle
Compounds are administered orally.
[0270] In another embodiment, the one or more Tricyclic Heterocycle
Compounds are administered intravenously.
[0271] In one embodiment, a pharmaceutical preparation comprising
at least one
[0272] Tricyclic Heterocycle Compound is in unit dosage form. In
such form, the preparation is subdivided into unit doses containing
effective amounts of the active components.
[0273] Compositions may be prepared according to conventional
mixing, granulating or coating methods, respectively, and the
present compositions can contain, in one embodiment, from about
0.1% to about 99% of the Tricyclic Heterocycle Compound(s) by
weight or volume. In various embodiments, the present compositions
can contain, in one embodiment, from about 1% to about 70% or from
about 5% to about 60% of the Tricyclic Heterocycle Compound(s) by
weight or volume.
[0274] The compounds of Formula I may be administered orally in a
dosage range of 0.001 to 1000 mg/kg of mammal (e.g., human) body
weight per day in a single dose or in divided doses. One dosage
range is 0.01 to 500 mg/kg body weight per day orally in a single
dose or in divided doses. Another dosage range is 0.1 to 100 mg/kg
body weight per day orally in single or divided doses. For oral
administration, the compositions may be provided in the form of
tablets or capsules containing 1.0 to 500 milligrams of the active
ingredient, particularly 1, 5, 10, 15, 20, 25, 50, 75, 100, 150,
200, 250, 300, 400, and 500 milligrams of the active ingredient for
the symptomatic adjustment of the dosage to the subject to be
treated. The specific dose level and frequency of dosage for any
particular subject may be varied and will depend upon a variety of
factors including the activity of the specific compound employed,
the metabolic stability and length of action of that compound, the
age, body weight, general health, sex, diet, mode and time of
administration, rate of excretion, drug combination, the severity
of the particular condition, and the host undergoing therapy.
[0275] For convenience, the total daily dosage may be divided and
administered in portions during the day if desired. In one
embodiment, the daily dosage is administered in one portion. In
another embodiment, the total daily dosage is administered in two
divided doses over a 24 hour period. In another embodiment, the
total daily dosage is administered in three divided doses over a 24
hour period. In still another embodiment, the total daily dosage is
administered in four divided doses over a 24 hour period.
[0276] The unit dosages of the Tricyclic Heterocycle Compounds may
be administered at varying frequencies. In one embodiment, a unit
dosage of a Tricyclic Heterocycle Compound may be administered once
daily. In another embodiment, a unit dosage of a Tricyclic
Heterocycle Compound may be administered twice weekly. In another
embodiment, a unit dosage of a Tricyclic Heterocycle Compound may
be administered once weekly. In still another embodiment, a unit
dosage of a Tricyclic Heterocycle Compound may be administered once
biweekly. In another embodiment, a unit dosage of a Tricyclic
Heterocycle Compound may be administered once monthly. In yet
another embodiment, a unit dosage of a Tricyclic Heterocycle
Compound may be administered once bimonthly. In another embodiment,
a unit dosage of a Tricyclic Heterocycle Compound may be
administered once every 3 months. In a further embodiment, a unit
dosage of a Tricyclic Heterocycle Compound may be administered once
every 6 months. In another embodiment, a unit dosage of a Tricyclic
Heterocycle Compound may be administered once yearly.
[0277] The amount and frequency of administration of the Tricyclic
Heterocycle Compounds will be regulated according to the judgment
of the attending clinician considering such factors as age,
condition and size of the subject as well as severity of the
symptoms being treated. The compositions of the invention can
further comprise one or more additional therapeutic agents,
selected from those listed above herein.
Kits
[0278] In one aspect, the present invention provides a kit
comprising a therapeutically effective amount of at least one
Tricyclic Heterocycle Compound, or a pharmaceutically acceptable
salt or prodrug of said compound and a pharmaceutically acceptable
carrier, vehicle or diluent.
[0279] In another aspect the present invention provides a kit
comprising an amount of at least one Tricyclic Heterocycle
Compound, or a pharmaceutically acceptable salt or prodrug of said
compound and an amount of at least one additional therapeutic agent
listed above, wherein the amounts of the two or more active
ingredients result in a desired therapeutic effect. In one
embodiment, the one or more Tricyclic Heterocycle Compounds and the
one or more additional therapeutic agents are provided in the same
container. In one embodiment, the one or more Tricyclic Heterocycle
Compounds and the one or more additional therapeutic agents are
provided in separate containers.
[0280] The present invention is not to be limited by the specific
embodiments disclosed in the examples that are intended as
illustrations of a few aspects of the invention and any embodiments
that are functionally equivalent are within the scope of this
invention. Indeed, various modifications of the invention in
addition to those shown and described herein will become apparent
to those skilled in the art and are intended to fall within the
scope of the appended claims.
[0281] A number of references have been cited herein, the entire
disclosures of which are incorporated herein by reference.
* * * * *